JP5703551B2 - Method for producing succinic acid - Google Patents

Description

  The present invention relates to succinic acid and a method for producing the same. In particular, the present invention relates to high-purity succinic acid purified from a fermentation broth obtained by microbial conversion such as glucose, sucrose, and cellulose derived from biomass resources, and a method for producing the same.

  Aliphatic dicarboxylic acids such as succinic acid and adipic acid are widely used as raw materials for polymers such as polyester and polyamide, and as synthetic raw materials for foods, pharmaceuticals and other chemicals. Among these uses, particularly when a dicarboxylic acid is used as a polymer raw material, a high-purity dicarboxylic acid is required to maintain the polymerization activity of the polymer and to obtain a high-quality polymer with little coloring. Moreover, when using succinic acid as a food material, it is required that the succinic acid produced does not contain odor components that impair the flavor.

  These aliphatic dicarboxylic acids have traditionally been produced industrially from petroleum-derived raw materials, but there are concerns about the recent depletion of fossil fuel resources and the background of global environmental problems such as an increase in carbon dioxide in the atmosphere. Therefore, a technique for deriving these polymer raw materials from biomass resources has attracted attention. These methods are expected to become a particularly important process from the viewpoint of carbon neutral since this resource is a renewable resource. In recent years, various aliphatic dicarboxylic acids can be produced from biomass resources with a very high carbon yield by fermentation using microorganisms.

  In the production of aliphatic dicarboxylic acids by fermentation, the raw material is generally a saccharide, such as glucose, sucrose, or cellulose. However, in these production methods of aliphatic dicarboxylic acids, there are cases where polysaccharides are contained as impurities in the aliphatic dicarboxylic acids, or that the raw material saccharides are not completely assimilated by microorganisms and remain mixed. . In addition, in the fermentation using microorganisms, when ammonia is used as a neutralizing agent, amino acids are also produced as impurities, and it has been difficult to remove these from aliphatic dicarboxylic acids by an efficient and inexpensive method.

  As a method for purifying an aliphatic dicarboxylic acid produced by fermentation, for example, a method using an ion exchange resin (for example, Patent Document 1) and a method using electrodialysis (for example, Patent Document 2) are known. In addition, as a salt exchange method, a method of producing a dicarboxylic acid and ammonium sulfate by mixing and reacting ammonium salt of dicarboxylic acid with ammonium hydrogen sulfate and / or sulfuric acid at a sufficiently low pH has been proposed (for example, Patent Document 3).

  However, even in the aliphatic dicarboxylic acid produced by these processes, when succinic acid is produced as the aliphatic dicarboxylic acid, an odor component that impairs flavor remains as a food material, or the polymer to be produced is a polymer raw material. It was still not a sufficiently pure dicarboxylic acid because it had a tendency to color. In addition, in the conversion reaction to a dicarboxylic acid derivative such as hydrogenation of an aliphatic dicarboxylic acid with a reduction catalyst to convert it to a diol compound, such a low-purity dicarboxylic acid is included in the dicarboxylic acid. Issues such as catalyst poisoning due to impurities remain.

US Pat. No. 6,284,904 JP-A-2-283289 Special table 2001-514900 gazette

  The present invention is an aliphatic dicarboxylic acid produced from a fermentation broth obtained by microbial conversion of glucose, sucrose, cellulose and the like derived from biomass resources, and as a raw material for food materials, polymers and dicarboxylic acid derivatives It is an object of the present invention to provide a succinic acid that is a useful high-purity aliphatic dicarboxylic acid and a method for producing the same.

  The present inventor has intensively studied to solve the above problems. As a result, the present inventors have found that succinic acid, which is an aliphatic dicarboxylic acid obtained by a fermentation method using microorganisms, is used as a food material, and a high-purity succinic acid useful as a raw material for polymers and dicarboxylic acid derivatives. In order to achieve this, it has been found that a method of hydrotreating a solution containing the succinic acid in the presence of a catalyst is effective, and particularly when used as a raw material for a polymer, in the ultraviolet region. The present inventors have obtained the knowledge that it is useful to reduce impurities that are absorbed in a specific region to a specific amount or less, thereby completing the present invention.

That is, the present invention is as follows.
(1) A method for producing succinic acid derived from biomass resources, comprising a step of hydrotreating at least a solution containing the succinic acid in the presence of a catalyst.
(2) The method for producing succinic acid according to (1), wherein the hydrogen treatment temperature is 30 ° C. or higher and 150 ° C. or lower, and the hydrogen pressure is 0.1 MPa or higher and 5 MPa or lower.

(3) The agglutination according to either (1) or (2), wherein hydrogen treatment is performed with a hydrogenation catalyst in the presence of any adsorbent selected from the group consisting of metal oxide, silica and activated carbon Acid production method.
(4) The method for producing succinic acid according to any one of (1) to (3), wherein the solution containing succinic acid is an aqueous solution.

(5) The method for producing succinic acid according to any one of (1) to (4), wherein fumaric acid is contained in a solution containing succinic acid to be subjected to hydrogen treatment.
(6) The production of succinic acid according to (5), wherein the content of fumaric acid in the solution containing succinic acid subjected to hydrogen treatment is 0.01 to 10% by weight with respect to the weight of succinic acid. Method.
(7) Any one of (1) to (6), wherein impurities are removed from a solution containing succinic acid so that an average absorbance in the ultraviolet region of 250 to 300 nm of succinic acid is 0.05 or less. A method for producing succinic acid according to claim 1.

(8) The method for producing succinic acid according to (7), wherein the impurities are removed by adsorption removal using activated carbon.
(9) The method for producing succinic acid according to (7) or (8), wherein the impurities are removed by crystallization using a solvent.
(10) The method for producing succinic acid according to any one of (1) to (9), wherein insoluble components in the solution containing succinic acid are removed by membrane permeation treatment in a step before the hydrogen treatment step. .

(11) The method for producing succinic acid according to any one of (1) to (9), wherein an insoluble component in the solution containing succinic acid is removed by an adsorbent in a step before the hydrogen treatment step.
(12) A succinic acid derived from biomass resources is subjected to a hydrogen treatment in the presence of a catalyst after at least one of an adsorption treatment using activated carbon in a solution and a crystallization treatment. The manufacturing method of the succinic acid in any one of (1)-(11).

(13) The method for producing succinic acid according to any one of (1) to (12), wherein the biomass resource is a plant resource.
(14) The method for producing succinic acid according to any one of (1) to (13), wherein the yellowness (YI) of the succinic acid is 10 or less.
(15) The method for producing succinic acid according to any one of (7) to (9), wherein the impurity is a compound containing nitrogen.

(16) The method for producing succinic acid according to any one of (7) to (9), wherein the impurity is an aromatic compound .

  According to the method of the present invention, succinic acid, which is a high-purity aliphatic dicarboxylic acid that is a raw material for a polymer that is not only colored as a food material but has little coloration and excellent mechanical properties and can be used in various applications, and As a raw material for the dicarboxylic acid derivative, it is possible to provide succinic acid which is a high-purity aliphatic dicarboxylic acid with little catalyst poisoning during the conversion reaction. The development of the present invention can greatly contribute to the solution of environmental problems, fossil fuel resource depletion problems, and the like.

The figure which shows the construction procedure of plasmid pMJPC17.2. The underlined number indicates a primer consisting of the sequence of SEQ ID NO. It is a conceptual diagram of a multistage extraction method. It is a figure which shows the absorption spectrum of the succinic acid obtained in the Example. It is a figure which shows the absorption spectrum of the succinic acid obtained by the comparative example.

Hereinafter, the present invention will be described in detail.
The present invention is a method for producing succinic acid derived from biomass resources, and the production process includes a step of hydrotreating at least a solution containing the succinic acid in the presence of a catalyst. The present invention relates to a method for producing succinic acid. In addition, succinic acid derived from biomass resources characterized in that the average absorbance in the ultraviolet region of 250 to 300 nm is 0.05 or less, and a method for producing succinic acid, wherein succinic acid is derived from biomass resources Further, the present invention relates to a method for producing succinic acid, wherein impurities are removed from the succinic acid so that an average absorbance in the ultraviolet region of 250 to 300 nm of the succinic acid is 0.05 or less.

<Aliphatic dicarboxylic acid>
The succinic acid of the present invention particularly relates to succinic acid that can be derived from biomass resources with a very high carbon yield.

  In the present invention, these succinic acids are derived from biomass resources. Biomass resources include, for example, wood, rice straw, rice husk, rice bran, old rice, corn, sugar cane, cassava, sago palm, okara, corn cob, tapioca cass, bagasse, vegetable oil scum, persimmon, buckwheat, soybean, fat, waste paper, papermaking residue , Marine product residue, livestock excrement, sewage sludge, food waste and the like. Among them, 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, buckwheat, soy, oil, waste paper, paper residue More preferred are wood, rice straw, rice husk, old rice, corn, sugar cane, cassava, sago palm, straw, oil and fat, waste paper, papermaking residue, and most preferred are corn, sugar cane, cassava, sago palm. These biomass resources generally contain a large amount of alkali metals and alkaline earth metals such as nitrogen element, Na, K, Mg, and Ca.

  These biomass resources are not particularly limited, but are induced to carbon sources through known pretreatment and saccharification processes such as chemical treatment with acids and alkalis, biological treatment with microorganisms, physical treatment, and the like. Is done. The process is not particularly limited, but includes, for example, a refinement process by pretreatment such as chipping, scraping, or crushing biomass resources. If necessary, a grinding step with a grinder or a mill is further included. The biomass resources thus refined are further guided to a carbon source through pretreatment and saccharification processes. Specific methods include acid treatment with strong acids such as sulfuric acid, nitric acid, hydrochloric acid, and phosphoric acid, and alkali treatment. Chemical methods such as treatment, ammonia freezing steam explosion method, solvent extraction, supercritical fluid treatment, oxidizing agent treatment, physical methods such as fine grinding, steam explosion method, microwave treatment, electron beam irradiation, microorganism and enzyme treatment Biological treatments such as hydrolysis by

  Carbon sources derived from the above biomass resources usually include hexoses such as glucose, mannose, galactose, fructose, sorbose and tagatose, pentoses such as arabinose, xylose, ribose, xylulose and ribulose, maltose, sucrose, lactose and trehalose. , Starch, cellulose and other disaccharides / polysaccharides, 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 acid, lignoceric acid, ceracolic acid, glycerin, mannitol, xylitol, ribitol, etc. Fermentable carbohydrates such as polyalcohols are used, and among these, hexoses such as glucose, mannose, galactose, fructose, sorbose, tagatose, pentoses such as arabinose, xylose, ribose, xylulose, ribulose, maltose, sucrose, Disaccharides and polysaccharides such as lactose, trehalose, starch, and cellulose are preferable, and among these, glucose, maltose, fructose, sucrose, lactose, trehalose, and cellulose are preferable.

  In the present invention, succinic acid is derived from a carbon source derived from these biomass resources. Specifically, for example, using these carbon sources, fermentation methods by microbial conversion, chemical conversion methods including reaction steps such as hydrolysis, dehydration reaction, hydration reaction, oxidation reaction, reduction reaction, and these fermentation methods Among them, succinic acid is synthesized by a combination of chemical conversion methods, and among these, fermentation methods based on microbial conversion using microorganisms having succinic acid-producing ability are preferable.

Microorganisms capable of producing succinic acid are not particularly limited as long as they are microorganisms capable of producing succinic acid, but include intestinal bacteria such as Escherichia coli, Bacillus bacteria, coryneform bacteria, aerobic microorganisms, facultative It is preferable to use anaerobic microorganisms or microaerobic microorganisms.
Examples of aerobic microorganisms include Coryneform Bacterium, Bacillus genus bacteria, Rhizobium genus bacteria, Arthrobacter genus bacteria, Mycobacterium genus bacteria, Rhodococcus (cc) Examples include genus bacteria, Nocardia bacteria, and Streptomyces bacteria, and coryneform bacteria are more preferable.

  The coryneform bacterium is not particularly limited as long as it is classified as such, and examples thereof include bacteria belonging to the genus Corynebacterium, bacteria belonging to the genus Brevibacterium, and bacteria belonging to the genus Arthrobacter. Examples include those belonging to the genus Corynebacterium or Brevibacterium, more preferably, Corynebacterium glutamicum, Brevibacterium flavum, Brevibacterium ammoniagenes (Brevibaterium). bacteria classified as Ammoniagenes) or Brevibacterium lactofermentum It is below.

As the microorganism having succinic acid-producing ability, it is preferable to use a strain having enhanced pyruvate carboxylase activity and reduced lactate dehydrogenase activity, as described in Examples below.
Reaction conditions such as reaction temperature and pressure in microbial conversion will depend on the activity of microorganisms such as selected cells and fungi, but suitable conditions for obtaining succinic acid should be selected in each case. That's fine.

  In microbial conversion, neutralizing agents are usually used because the metabolic activity of microorganisms decreases or the activity of microorganisms stops when pH decreases, and the production yield deteriorates or the microorganisms die. To do. Usually, the pH in the reaction system is measured by a pH sensor, and the pH is adjusted by adding a neutralizing agent so as to be in a predetermined pH range. The pH value is adjusted to a range where the activity is most effectively exhibited according to the type of microorganisms such as fungus bodies and molds to be used. There is no restriction | limiting in particular about the addition method of 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. Ammonia, ammonium carbonate and urea are preferred. Examples of the alkali (earth) metal hydroxide include NaOH, KOH, Ca (OH) ₂ , Mg (OH) ₂ , or a mixture thereof. Examples of the alkali (earth) metal carbonate include _{is, Na 2 CO 3, K 2} CO 3, CaCO 3, MgCO 3, NaKCO 3 etc., or the like and mixtures thereof.

  In the present invention, the fermentation broth after microbial conversion may be appropriately concentrated in consideration of operability and efficiency in the subsequent purification step. Although it does not specifically limit as a concentration method, The method of distribute | circulating an inert gas, the method of distilling off water by heating, the method of distilling off water by pressure reduction, the method of combining these, etc. are mentioned. Further, the concentration operation may be performed by a batch operation or a continuous operation.

In addition, 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 the microorganisms is not particularly limited, and sedimentation separation, centrifugation, filtration separation, a combination thereof, and the like are used. Industrially, it is performed by a method such as centrifugal separation or membrane filtration separation. In centrifugation, centrifugal sedimentation, centrifugal filtration, or the like can be used. In the centrifugation, the operating conditions are not particularly limited, but the separation is usually performed with a centrifugal force of 100 G to 100,000 G. The operation can be performed continuously or batchwise.

  In membrane filtration separation, microfiltration and / or ultrafiltration can be used. The material of the film is not particularly limited, and for example, an organic film such as polyolefin, polysulfine, polyacrylonitrile, polyvinylidene fluoride, or a film made of an inorganic material such as ceramic can be used. As an operation method, either a dead end type or a cross flow type can be used. In membrane filtration separation, since microorganisms often clog the membrane, a method of performing membrane filtration after roughly removing the microorganisms by centrifugation or the like is also used.

  In the present invention, when a neutralizing agent is used as described above, a succinic acid salt is obtained, and the obtained succinic acid salt is converted into the desired succinic acid. The succinic acid may be converted to succinic acid by a reaction crystallization method using a weakly acidic organic acid having a higher acid dissociation constant (pKa) than the target succinic acid. An example of such an organic acid is acetic acid. Alternatively, the succinic acid salt obtained as described above may be converted to succinic acid with an inorganic acid. Here, examples of the inorganic acid used include sulfuric acid, hydrochloric acid, carbonic acid, phosphoric acid, and nitric acid. More specifically, in the case of succinic acid fermentation, when fermentation is performed while neutralizing the produced succinic acid with ammonia or magnesium hydroxide, ammonium succinate or magnesium succinate is produced in the fermentation broth. When such a fermented solution containing ammonium succinate or magnesium succinate is treated with sulfuric acid or the like, an aqueous solution containing succinic acid is obtained.

  In the present invention, the solution or aqueous solution containing succinic acid is a solution or aqueous solution mainly containing succinic acid derived from biomass resources. Accordingly, a solution or an aqueous solution mainly containing a succinate such as ammonium succinate or magnesium succinate is referred to as a solution or aqueous solution containing succinate. The term “contained as the main component” as used herein means that the weight of the component relative to the weight of all components excluding the solvent is usually 50% by weight or more, preferably 60% by weight or more, more preferably 70% by weight or more, and particularly preferably The state of 90% by weight or more is shown.

In the present invention, the aqueous solution converted into succinic acid with the inorganic acid is not particularly limited, but succinic acid may be extracted from the aqueous solution using an organic solvent.
The organic solvent used in this method usually has an inorganic value / organic value ratio (I / O value) of 0.2 or more and 2.3 or less, and has a boiling point of 40 ° C. at normal pressure (1 atm). More preferably, the organic solvent is an organic solvent having an I / O value of 0.3 or more and 2.0 or less and a boiling point of 40 ° C. or more at normal pressure, and more preferably I / O. An organic solvent having a value of 0.3 or more and 2.0 or less and a normal pressure and a boiling point of 60 ° C. or more. By using such an organic solvent, succinic acid can be selectively extracted and efficiently separated from saccharides and amino acids. In addition, by using an organic solvent having a boiling point of 40 ° C. or higher at normal pressure, there is a risk that the solvent vaporizes and ignites, a problem that the solvent vaporizes and the extraction efficiency of succinic acid decreases, and the solvent is recycled. The problem of difficulty can be avoided.

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.
Organic solvents 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, and ether solvents such as tetrahydrofuran and dioxane. Examples include 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.

The I / O value and boiling point of each solvent are shown below.
I O I / O Boiling point tetrahydrofuran 30 80 0.375 66.0
Methyl ethyl ketone 65 60 1.083 79.6
Methyl isobutyl ketone 65 120 0.542 94.2
Acetone 65 40 1.625 56.1
Acetonitrile 70 40 1.750 81.1
Ethyl acetate 85 80 1.063 77.2
Propanol 100 60 1.667 97.2
Isobutanol 100 70 1.429 108.0
Octanol 100 160 0.625 179.8
Dioxane 40 80 0.500 101.3
In the extraction step, the organic solvent is preferably added in a volume of 0.5 to 5 with respect to the volume 1 of the aqueous solution containing succinic acid, and in a volume of 1 to 3 with respect to the volume 1 of the aqueous solution containing succinic acid. More preferably, it is added.

Although the temperature of an extraction process should just be a temperature from which succinic acid is extracted, 10-90 degreeC is preferable and 20-85 degreeC is more preferable.
Through the extraction step, succinic acid is recovered in an organic solvent, and impurities such as ammonia, sulfur-containing impurities, metal cations, and the like derived from fermentation bacteria are separated to some extent in addition to saccharides and nitrogen elements derived from fermentation. In addition, in order to extract succinic acid more efficiently, the extraction process with an organic solvent may be repeated a plurality of times, or countercurrent extraction may be performed.

  In the present invention, it is possible to reduce the amount of many impurities such as nitrogen element derived from fermentation bacteria, ammonia, sulfur-containing impurities and metal cations in addition to nitrogen element contained in biomass resources by purification from fermentation broth containing succinic acid. Although important, especially reduce the content of odorous components contained in succinic acid, or reduce the amount of impurities that absorb in the ultraviolet region of 250-300 nm until the average absorbance is 0.05 or less. Is essential.

  Known methods for removing odor components include a deodorizing method using an adsorbent such as activated carbon, a washing and removing method using an organic solvent, a crystallization method, and an aeration method. In the present invention, it has surprisingly been found that hydrotreating in the presence of a catalyst is particularly effective for removing odorous components. On the other hand, a small amount of fumaric acid may be contained in the succinic acid-containing liquid derived from biomass resources by fermentation or the like.

  When hydrogen treatment of a succinic acid-containing liquid derived from biomass resources by fermentation or the like as in the present invention is performed, not only odor components in succinic acid can be easily removed, but fumaric acid is contained as described above. In this case, succinic acid is produced from fumaric acid, and the yield improvement of succinic acid is achieved at the same time. Therefore, the hydrotreating method is a particularly excellent technique that has not been conventionally used as a deodorizing method for succinic acid. Therefore, in the present invention, hydrotreating of a solution containing succinic acid derived from fermentation in the presence of a catalyst is an important step in the purification process.

Further, in the present invention, in order to reduce impurities having absorption in the ultraviolet region of 250 to 300 nm contained in this succinic acid to an average absorbance of 0.05 or less, it is usually produced as described above. The succinic acid must be subjected to hydrogen treatment and further combined with purification treatment such as crystallization treatment and activated carbon treatment.
The hydrogen treatment in the present invention may be either a batch type or a continuous type, and can be performed according to a conventionally known method. As a specific method of hydrogen treatment, a solution containing succinic acid and a hydrogenation catalyst are allowed to coexist in a pressurized reactor, hydrogen gas is introduced while stirring the mixture, and hydrogen treatment is performed. A method in which a reaction solution containing succinic acid is separated from a hydrogenation catalyst and taken out from the reactor, and a succinic acid-containing solution and hydrogen gas are circulated from the lower part of the reactor using a fixed-bed multitubular or single tube reactor. Performing hydrogen treatment and removing the treated succinic acid-containing reaction solution or hydrogen gas from the lower part of the reactor and the succinic acid-containing solution from the upper part to perform the hydrogen treatment, and treating the treated succinic acid-containing reaction liquid. The method of taking out is mentioned.

As the hydrogenation catalyst, known homogeneous and heterogeneous noble metal-containing hydrogenation catalysts can be used. Specific examples include, but are not limited to, hydrogenation catalysts containing noble metals such as ruthenium, rhodium, palladium and platinum. Among these, hydrogenation catalysts containing palladium and platinum, particularly palladium, are preferred.
These hydrogenation catalysts can be used with the above compounds containing noble metals as they are or in the presence of a ligand such as an organic phosphine, but heterogeneous noble metal-containing catalysts for reasons of ease of catalyst separation. Is preferred.

  In addition, these noble metal-containing compounds can be subjected to hydrogen treatment in the presence of a metal oxide such as silica, titanium, zirconia, or activated alumina, or a composite metal oxide thereof, or activated carbon. This method is a preferred embodiment because not only the odor components contained in the succinic acid derived from fermentation, but also the colored components and organic impurities can be adsorbed and removed simultaneously, and the impurities can be removed efficiently. The same effect is also achieved when using a catalyst in which the above-mentioned noble metal is supported on a metal oxide such as silica, titanium, zirconia, activated alumina or a composite metal oxide thereof, or a support such as activated carbon. A method using these supported catalysts is also preferably used. The supported amount of the noble metal is usually 0.1 to 10% by weight of the support, but the support is not particularly limited, but silica or activated carbon, particularly activated carbon is used because of the small amount of metal elution during the hydrogen treatment. preferable.

  Therefore, in the present invention, the embodiment in which the hydrogen treatment is performed with a hydrogenation catalyst in which a noble metal is supported on a metal oxide such as silica, titanium, zirconia, activated alumina, or a composite metal oxide thereof, or a support such as activated carbon is the present invention. It is included in the definition of the embodiment in which the hydrogen treatment is performed with the hydrogenation catalyst in the presence of any adsorbent selected from the group of metal oxide, silica and activated carbon.

  Solvents containing succinic acid derived from biomass resources during hydrogen treatment are water, organic acids such as acetic acid and propionic acid, esters such as ethyl acetate, methanol, ethanol, propanol, isopropanol, butanol, 2-ethyl Alcohols such as -1-hexanol and isobutanol, ethers such as diethyl ether, di-n-butyl ether, diisopropyl ether, di-n-butyl ether, tetrahydrofuran and dioxane, ketones such as acetone, methyl ethyl ketone and diethyl ketone, acetonitrile Nitriles such as these, and mixed solvents thereof can be used, but water is most preferred among them. Usually, deionized water, distilled water, river water, well water, tap water, etc. are used as the water, but if necessary, succinic acid is crystallized from the succinic acid-containing reaction solution in the subsequent step after the hydrogenation reaction. The solution after filtration can be used repeatedly. The succinic acid concentration in the solution may be equal to or lower than the saturation solubility at the liquid temperature.

  The content of fumaric acid contained in the succinic acid subjected to hydrotreating is usually 0.01% by weight or more, preferably 0.05% by weight or more, and the upper limit is 10% by weight, based on the weight of succinic acid. Hereinafter, it is preferably 5% by weight or less. If the content of fumaric acid is too low, the purification process up to the hydrotreating step becomes complicated. On the other hand, if the content of fumaric acid is too high, fumaric acid has a long time for hydrotreating or has low solubility. This causes a problem that the succinic acid solution having a high concentration cannot be prepared.

Hydrogen used may be pure hydrogen, but hydrogen diluted with an inert gas such as nitrogen, helium or argon can also be used. The carbon monoxide concentration in the hydrogen gas is usually 10000 ppm or less, preferably 2000 ppm or less, more preferably 1000 ppm or less because there is concern about the influence on the hydrogen treatment efficiency.
The lower limit of the hydrogen pressure during the hydrogen treatment is usually 0.1 MPa or more, and the upper limit is usually 5 MPa or less, preferably 3 MPa or less, more preferably 1 MPa or less. If the hydrogen pressure is too low, the reaction rate is slow and it takes time to complete the reaction. On the other hand, if it is too high, succinic acid hydrides such as butanediol and tetrahydrofuran may be produced as a by-product depending on the catalyst and reaction conditions. It is not preferable.

The lower limit of the temperature during the hydrogen treatment is usually 30 ° C or higher, preferably 50 ° C or higher, and the upper limit is usually 150 ° C or lower, preferably 120 ° C or lower. If the reaction temperature is too low, the reaction rate is slow and it takes time to complete the reaction.If the reaction temperature is too high, by-products such as succinic acid hydride and when using water as a solvent, by-products such as malic acid are not produced. It is not preferable because it increases.
As the crystallization solvent in the crystallization treatment in the present invention, water, organic acids such as acetic acid and propionic acid, esters such as ethyl acetate, methanol, ethanol, propanol, isopropanol, butanol, 2-ethyl-1-hexanol, iso Alcohols such as butanol, diethyl ether, di-n-butyl ether, diisopropyl ether, di-n-butyl ether, ethers such as tetrahydrofuran, dioxane, ketones such as acetone, methyl ethyl ketone, diethyl ketone, nitriles such as acetonitrile, and Although these mixed solvents can be used, water is most preferable among these. Usually, deionized water, distilled water, river water, well water, tap water, etc. are used for water.

  The crystallization temperature can be selected from the range of about 0 to 90 ° C., preferably about 0 to 85 ° C., and the cooling rate can be selected from the range of about 1 to 120 ° C./hr, preferably about 5 to 60 ° C./hr. It is performed under reduced pressure (for example, about 1 atm), reduced pressure or increased pressure. The aging time can be appropriately selected from a range of about 0.1 to 5 hours, preferably 0.5 to 4 hours, and more preferably about 0.5 to 2 hours.

Further, in the present invention, the average absorbance is 0.05 or less for the amount of impurities that absorb in the ultraviolet region of 250 to 300 nm contained in succinic acid by combining the above crystallization treatment and, if necessary, activated carbon treatment. It is preferable to reduce it until it becomes.
As the activated carbon used in the present invention, any known ones such as coal-based, wood-based, coconut-based, and resin-based can be used. Also, use activated carbon activated by various methods such as gas activation method, water vapor activation method, chemical activation method such as zinc chloride and phosphoric acid, etc. Can do.

  Specifically, Calgon CPG, Calgon CAL, Calgon SGL, Diasorb W, Diamond Hope MS10, Diamond Hope M010, Diamond Hope MS16, Diamond Hope 6MD, Diamond Hope 6MW, Diamond Hope 8ED, Diamond Hope ZGN4, Centur, Japan Norit Stock, manufactured by Mitsubishi Chemical Calgon Co., Ltd. Company-made GAC, GAC PLUS, GCN PLUS, C GRAN, RO, ROX, DARCO, CN, SX, SX PLUS, sa, SX, PK, W, Kuraray Chemical Co., Ltd. GW, GWH, GLC, 4GC, KW , PW, PK, HC-30S, GL-30S, 4G-3S, PA, PC, manufactured by Tsurumi Co., Ltd., P, W, CW, SG, SGP, S, GB, CA, K, manufactured by Futamura Chemical Co., Ltd. Hakuho KL, Hakuho W2c, Hakuho WH2c, Hakuho W5c, Hakuho WH5X, Hakuho XS7100H-3, Calaburafin, Hakuho A, Hakuho C, Hakuho M, Ajinomoto Fine Techno Co., Ltd. Hokuetsu CL-K, Hokuetsu Hs, Hokuetsu KS, etc. Is mentioned.

Among these, coconut charcoal and wood charcoal are preferred because impurities that absorb in the ultraviolet region of 250 to 300 nm contained in the aliphatic dicarboxylic acid can be efficiently removed. On the other hand, from the viewpoint of efficiently removing the coloring component of succinic acid, activated carbon activated by a method such as a gas activation method, a water vapor activation method, a chemical activation method such as zinc chloride or phosphoric acid is preferred, and among them, the water vapor activation method, Chemical activated carbon such as zinc chloride or phosphoric acid is preferred, and activated carbon activated by chemicals such as zinc chloride or phosphoric acid is particularly preferred. The shape of the activated carbon used may be any of powdered coal, crushed coal, formed coal, and fibrous activated carbon. When used in a column, granular and granular activated carbon is preferred for the purpose of suppressing the tower pressure. The activated carbon treatment method is a batch method of mixing with activated carbon followed by filtration and separation, and passing through a packed bed of activated carbon. Either method of liquefaction is possible. The processing time is usually 5 minutes to 5 hours, preferably 10 minutes to 2 hours in the case of a batch system, and is usually 0.1 to 20 hr ⁻¹ as SV (space velocity) in the case of a packed bed system. The treatment temperature is usually 20 to 90 ° C.

Furthermore, in the present invention, for the purpose of removing impurities in succinic acid, a purification operation such as ion exchange column treatment may be used in combination.
Here, the ion exchange column treatment is to remove ions by passing a liquid to be treated through a column filled with an ion exchange resin. The ion exchange resin should be selected according to the ions contained in the liquid to be treated and the required purity of succinic acid. For example, anion exchange resins are used to remove anions such as sulfate ions and chloride ions. In order to remove cations such as metal ions and ammonium ions from (OH type), a cation exchange resin (H type) can be used, but both of them may be used if necessary.

  Ion exchange resins are classified into strongly acidic cation exchange resins, weakly acidic cation exchange resins, strongly basic anion exchange resins, and weakly basic anion exchange resins depending on the strength of the functional group as an acid or base. Although it is classified into a gel type and a porous type, the ion exchange resin used here is not particularly limited. However, in consideration of the efficiency of ion exchange, it is preferable to use a strongly acidic cation exchange resin or a strongly basic anion exchange resin having a stronger strength as an acid or a base. Further, it is desirable to use a more versatile and inexpensive gel type without any special reason that the shape is also porous. Specifically, Diaion SK1B (H type) is exemplified as a cation exchange resin, and Diaion SA10A is exemplified as an anion exchange resin.

  The ion exchange column treatment can be performed within a temperature range that is equal to or higher than the temperature at which succinic acid is dissolved in the liquid to be treated and lower than the heat resistant temperature of the ion exchange resin. That is, with a cation exchange resin, the treatment is usually performed at 20 to 100 ° C., depending on the succinic acid concentration in the liquid to be treated. On the other hand, since anion exchange resins have lower heat resistance than cation exchange resins, treatment is usually performed at 10 to 80 ° C. In the case of using anion exchange column treatment from the viewpoint of treatment temperature, a step where the succinic acid concentration is low and the column treatment can be performed at a low temperature is desirable.

The liquid flow treatment method is not particularly limited, but the treatment is usually performed at a space velocity (SV) of 0.1 to 10 hr ⁻¹ and a superficial velocity of ^{1 to} 20 m / hr. If the processing speed is too high, pressure loss before and after the column becomes large, and ion exchange becomes insufficient. Conversely, if the processing speed is unnecessarily slow, the column becomes unnecessarily large.
Usually, in the column treatment, the ion concentration is measured regularly or periodically at the column outlet, and if ion leakage is observed at the column outlet, the ion exchange resin is regenerated. The regeneration of the ion exchange resin can be carried out in accordance with a normal method, using an acid such as sulfuric acid or hydrochloric acid for a cation exchange resin, or an alkali such as caustic soda for an anion exchange resin.

  In the present invention, when the obtained succinic acid is used as a polymer raw material, it is essential to reduce the amount of impurities that absorb in the ultraviolet region of 250 to 300 nm to an average absorbance of 0.05 or less. However, the average absorbance is preferably 0.03 or less, and particularly preferably 0.01 or less. As is apparent from this example, succinic acid having a high average absorbance is markedly colored when used as a raw material for polyester.

The absorbance in the present invention is a value measured with an ultraviolet-visible absorption spectrophotometer after placing a 3.0 wt% aqueous succinic acid solution in a quartz cell having an optical path length of 1 cm. In the present invention, the absorbance was measured using an ultraviolet-visible absorption spectrophotometer (Hitachi spectrophotometer UV-3500), but can be measured using a commercially available ultraviolet-visible absorption spectrophotometer.
Here, the absorbance (A) is the absorbance when measured at an optical path length of 1 cm, and is a value calculated according to the following definition.

A = log ₁₀ (I ₀ / I) (where I ₀ = incident light intensity and I = transmitted light intensity)
The average absorbance in the ultraviolet region of 250 to 300 nm is a value obtained by dividing the sum of absorbances measured every 1 nm between 250 and 300 nm by 51.
Average absorbance = (total absorbance every 1 nm between 250 and 300 nm) / 51
In the present invention, the impurities that absorb in the ultraviolet region of 250 to 300 nm are not particularly limited, and examples thereof include compounds having a nitrogen element and compounds having aromaticity. Examples of such compounds include oxygen-containing heteroaromatic compounds such as furan, nitrogen-containing heteroaromatic compounds such as pyrrole, pyridine, and pyrazine, and benzene aromatic compounds such as phenol, benzaldehyde, and benzoic acid. Specifically, furfural, furfuryl alcohol, methylfurfuryl alcohol, hydroxymethylfurfural, furosine, 2-pyrrolecarboxaldehyde, pyrrolecarboxylic acid, methylpyrrolecarboxylic acid, pyridinecarboxylic acid, pyridinedicarboxylic acid, methylpyridinecarboxylic acid, Methylpyridinedicarboxylic acid, pyrazine, 2-methylpyrazine, dimethylpyrazine, trimethylpyrazine, tetramethylpyrazine, phenol, benzoic acid, monohydroxybenzoic acid such as salicylic acid and creosote acid, dihydroxybenzoic acid such as pyrocatechuic acid and protocatechuic acid, Examples include trihydroxybenzoic acid such as gallic acid, aromatic aldehydes such as benzaldehyde, methylbenzaldehyde, dimethylbenzaldehyde, and mixtures thereof. That. In addition, although the said compound may contain an isomer, the illustration of the said compound shall include all the isomers.

In the present invention, since the impurity species to be removed differ depending on the type of activated carbon as described above, these impurity removal methods include a method of combining a plurality of activated carbon species, activated carbon treatment, and the above-described hydrogen treatment and crystallization treatment. The method of combining these is mentioned.
Moreover, when using water as a solvent, the insoluble component may be mixed in the succinic acid solution derived from fermentation. Such an insoluble component is a factor that reduces the efficiency of the above-described impurity removal by activated carbon and the subsequent purification process, so it is preferable to remove the insoluble component in advance. Insoluble components are removed from the succinate produced by the fermentation method to succinic acid, and the process from the succinic acid solution to the activated carbon treatment process is performed using a known membrane permeabilization process. Is preferred. As another method, the powdered activated carbon is allowed to coexist to adsorb insoluble components to improve the permeability of the membrane permeation treatment, or the above-mentioned impurities are simultaneously adsorbed and removed together with the insoluble components using appropriate powdered activated carbon. Are also preferably used.

Furthermore, in the present invention, when removing impurities by combining crystallization or / and activated carbon treatment and hydrogen treatment, although not particularly limited, since impurities are efficiently removed, A process in which the crystallization or / and activated carbon treatment step is performed before is preferably used.
The succinic acid recovered by crystallization depends on its use, but can be dried by a conventional method. Usually, the moisture content of succinic acid is 0.1-2% by weight, preferably 0.2-1% by weight. The drying method is not particularly limited, and a direct heating method in which heating is performed directly with warm air, an indirect heating method using steam, or the like can be used. For example, box dryers, band dryers, rotary dryers and the like are used as dryers using hot air, and drum dryers and disk dryers are used as dryers using indirect heating. The operating pressure may be normal pressure or reduced pressure. Furthermore, the operation method may be batch operation or continuous operation. As for the warm air temperature, the heating surface temperature is usually 20 to 200 ° C, preferably 50 to 150 ° C. If the temperature is too low, a high pressure reduction is required for drying. Conversely, if the temperature is too high, succinic acid is dehydrated and succinic anhydride is generated, which is not preferable.

  Moreover, it is preferable that the succinic acid manufactured by this invention is a thing with little absorption normally in the visible light region and little coloring. The upper limit of the yellowness (YI value) of the succinic acid of the present invention is usually 10 or less, preferably 6 or less, more preferably 4 or less, while the lower limit is not particularly limited. It is 10 or more, preferably -5 or more, more preferably -1 or more. Succinic acid exhibiting a high YI value has a drawback that when used as a polymer raw material, coloring of the produced polymer becomes remarkable. On the other hand, succinic acid exhibiting a low YI value is a more preferable form, but is economically disadvantageous in that it requires a very large amount of capital investment for its production and requires a large amount of production time. In the present invention, the YI value is a value measured by a method based on JIS K7105.

The succinic acid of the present invention may contain elemental nitrogen as an impurity due to a biomass resource-derived, fermentation treatment, and purification treatment including a neutralization step with an acid. Specifically, nitrogen elements such as amino acids, proteins, ammonium salts, urea, and fermenting bacteria may be included.
The nitrogen atom content contained in the aliphatic dicarboxylic acid of the present invention is usually 2000 ppm or less, preferably 1000 ppm or less, more preferably 100 ppm or less, most preferably 20 ppm or less in terms of atoms in dicarboxylic acid. is there. The lower limit is usually 0.01 ppm or more, preferably 0.05 ppm or more, more preferably 0.1 ppm or more, and still more preferably 1 ppm or more for reasons of economic efficiency of the purification step.

The nitrogen atom content is measured by a known method such as elemental analysis or an amino acid analyzer and a method of separating amino acids and ammonia in a sample under biological amino acid separation conditions and detecting them by ninhydrin color development. Value.
When succinic acid having a nitrogen atom content in the above range is used as a polyester raw material, it is advantageous in reducing the coloration of the resulting polyester. It also has the effect of suppressing the delay of the polyester polymerization reaction.

Further, the succinic acid of the present invention may contain sulfur atoms due to a purification treatment including a neutralization step with an acid. Specifically, examples of the impurities containing sulfur atoms include sulfuric acid, sulfate, sulfurous acid, organic sulfonic acid, and organic sulfonate.
The sulfur atom content contained in the succinic acid of the present invention is, in terms of atoms in dicarboxylic acid, the upper limit is usually 100 ppm or less, preferably 20 ppm or less, more preferably the upper limit is 10 ppm or less, and particularly preferably the upper limit. Is 5 ppm or less, and most preferably the upper limit is 0.5 ppm or less. On the other hand, the lower limit is usually 0.001 ppm or more, preferably 0.01 ppm or more, more preferably 0.05 ppm or more, and particularly preferably 0.1 ppm or more. When the amount is too large, when used as a polyester raw material, there is a tendency that the polymerization reaction is delayed, the resulting polymer is partially gelled, the stability of the produced polymer is lowered, and the like. On the other hand, an excessively small system is a preferred form, but the purification process becomes complicated and economically disadvantageous. The sulfur atom content is a value measured by a known elemental analysis method.

The succinic acid of the present invention may contain an alkali metal element. The content of the alkali metal contained in the aliphatic dicarboxylic acid is usually 50 ppm or less, preferably 30 ppm or less, more preferably 10 ppm or less, and particularly preferably 5 ppm or less. If the content is too high, when used as a polymer raw material, it not only reduces thermal stability and hydrolysis resistance, but also causes severe polymerization inhibition during polymerization and high polymerization with sufficient mechanical properties for practical use. Degree of polymer may not be obtained.
<Manufacture of polyester>
The polyester of the present invention contains succinic acid units and diol units as essential components. In the present invention, the succinic acid constituting the succinic acid unit is a dicarboxylic acid containing high-purity succinic acid derived from the biomass resource. Therefore, in the present invention, a mixture of an aliphatic and / or aromatic dicarboxylic acid derived from a fossil resource and succinic acid derived from the biomass resource is also preferably used.

<Diol unit>
In the present invention, the diol unit is derived from an aliphatic diol, and a known compound can be used.
The aliphatic diol is not particularly limited as long as it is an aliphatic and alicyclic compound having two OH groups, but the lower limit of the carbon number is 2 or more, and the upper limit is usually 10 or less, preferably 6 The following aliphatic diols are mentioned.

Specific examples of the aliphatic diol include, for example, ethylene glycol, 1,3-propylene glycol, neopentyl glycol, 1,6-hexamethylene glycol, decamethylene glycol, 1,4-butanediol, and 1,4- And cyclohexanedimethanol. These may be used alone or as a mixture of two or more.
Among these, ethylene glycol, 1,4-butanediol, 1,3-propylene glycol and 1,4-cyclohexanedimethanol are preferable, and among them, ethylene glycol and 1,4-butanediol, and these And a mixture mainly composed of 1,4-butanediol or 1,4-butanediol is particularly preferred. The main component here means that the component is usually at least 50 mol%, preferably at least 60 mol%, more preferably at least 70 mol%, particularly preferably at least 90 mol%, based on all diol units. Show.

Further, both terminal hydroxy polyethers may be used by mixing with the above aliphatic diol. As both-end hydroxy polyether, the lower limit is usually 4 or more, preferably 10 or more, and the upper limit is usually 1000 or less, preferably 200 or less, more preferably 100 or less.
Specific examples of both terminal hydroxy polyethers include diethylene glycol, triethylene glycol, polyethylene glycol, polypropylene glycol, polytetramethylene glycol, poly 1,3-propanediol, and poly 1,6-hexamethylene glycol. . In addition, a copolymerized polyether of polyethylene glycol and polypropylene glycol can be used. The use amount of these both terminal hydroxy polyethers is an amount calculated as a content in the polyester of usually 90% by weight or less, preferably 50% by weight or less, more preferably 30% by weight or less.

<Other copolymer components>
In the present invention, a copolymer component may be added to the polyester in addition to the diol component and the dicarboxylic acid component.
Specific examples of the copolymer component include a bifunctional oxycarboxylic acid, an unsaturated dicarboxylic acid, a trifunctional or higher polyhydric alcohol and a trifunctional or higher polyhydric carboxylic acid or an anhydride thereof for forming a crosslinked structure. And at least one polyfunctional compound selected from the group consisting of trifunctional or higher functional oxycarboxylic acids. When these copolymer components are added, the effect of significantly improving the polymerization rate during the production of the polyester is exhibited. Among these copolymer components, oxycarboxylic acids are preferably used because polyesters having a high degree of polymerization tend to be easily produced.

  Specific examples of the bifunctional oxycarboxylic acid include lactic acid, glycolic acid, hydroxybutyric acid, hydroxycaproic acid, 2-hydroxy-3,3-dimethylbutyric acid, 2-hydroxy-3-methylbutyric acid, and 2-hydroxyiso Examples thereof include caproic acid, and these may be esters of oxycarboxylic acid, lactones, or derivatives of oxycarboxylic acid polymers. These oxycarboxylic acids can be used alone or as a mixture of two or more. When optical isomers exist in these, any of D-form, L-form, and racemic form may be sufficient, and a form may be a solid, a liquid, or aqueous solution. Of these, readily available lactic acid or glycolic acid is particularly preferable. The form is preferably 30 to 95% because an aqueous solution can be easily obtained. In this case, the amount of oxycarboxylic acid used is usually 0.02 mol% or more, preferably 0.5 mol% or more, more preferably 1.0 mol% or more, with respect to the raw material monomer. The upper limit is usually 30 mol% or less, preferably 20 mol% or less, more preferably 10 mol% or less.

  Examples of the unsaturated dicarboxylic acid include itaconic acid, aconitic acid, fumaric acid, maleic acid and the like, and they can be used alone or as a mixture of two or more. Since the amount of unsaturated dicarboxylic acid used is a cause of gel generation, it is usually 5 mol% or less, preferably 0.5 mol% or less, more preferably based on all monomer units constituting the polyester. 0.05 mol% or less.

Specific examples of the trifunctional or higher polyhydric alcohol include glycerin, trimethylolpropane, pentaerythritol and the like, and they can be used alone or as a mixture of two or more.
Specific examples of the tri- or higher functional polycarboxylic acid or anhydride thereof include propanetricarboxylic acid, pyromellitic anhydride, benzophenonetetracarboxylic anhydride, cyclopentatetracarboxylic anhydride, and the like. It can also be used as a mixture of two or more.

Specific examples of the tri- or higher functional oxycarboxylic acid include malic acid, hydroxyglutaric acid, hydroxymethylglutaric acid, tartaric acid, citric acid, hydroxyisophthalic acid, hydroxyterephthalic acid, and the like. It can also be used as a mixture of In particular, malic acid, tartaric acid, and citric acid are preferable because of easy availability.
Since the amount of the trifunctional or higher functional compound is a cause of gel generation, it is usually 5 mol% or less, preferably 0.5 mol% or less, based on all monomer units constituting the polyester. More preferably, it is 0.2 mol% or less.

<Production method of polyester>
As a method for producing the polyester in the present invention, a conventionally known method can be used. For example, after performing an esterification reaction and / or a transesterification reaction between the aliphatic dicarboxylic acid component containing succinic acid and a diol component. It can also be produced by a general method of melt polymerization such as a polycondensation reaction under reduced pressure, or a known solution heating dehydration condensation method using an organic solvent. From the viewpoint, a method of producing a polyester by melt polymerization performed in the absence of a solvent is preferable.

Conventionally known ranges can be adopted for conditions such as temperature, time, and pressure.
The reaction temperature of the esterification reaction and / or transesterification reaction of the aliphatic dicarboxylic acid component containing succinic acid and / or the diol component is usually 150 ° C. or higher, preferably 180 ° C. or higher, and preferably 260 ° C. or lower, preferably upper limit. It is 250 degrees C or less. The reaction atmosphere is usually an inert gas atmosphere such as nitrogen or argon. The reaction pressure is usually normal pressure to 10 kPa, but normal pressure is preferred.

The reaction time is usually 1 hour or longer, and the upper limit is usually 10 hours or shorter, preferably 4 hours or shorter.
In the subsequent polycondensation reaction, the pressure is usually 0.01 × 10 ³ Pa or more at the lower limit, preferably 0.01 × 10 ³ Pa or more, and the upper limit is usually 1.4 × 10 ³ Pa or less, preferably 0.4. It is performed under a vacuum degree of × 10 ³ Pa or less. If the pressure at the time of polymerization production is too high, the polymerization production time of the polyester becomes long, and accordingly, molecular weight reduction and coloration due to thermal decomposition of the polyester are caused, and it tends to be difficult to produce a polyester having practically sufficient characteristics. . On the other hand, the method of producing using an ultra-high vacuum polymerization equipment is a preferred embodiment from the viewpoint of improving the polymerization rate, but it requires not only a very expensive equipment investment, but still requires a long polymerization production time for polyester. Since there is a tendency, there is a concern about molecular weight reduction or coloring due to thermal decomposition of polyester.

  The lower limit of the reaction temperature is usually 150 ° C or higher, preferably 180 ° C or higher, and the upper limit is usually 280 ° C or lower, preferably 260 ° C or lower. If this temperature is too low, particularly in the present invention, the polymerization reaction rate becomes extremely slow, and not only does it take a long time to produce a polyester with a high degree of polymerization, but also requires a high-powered stirrer, which is economically disadvantageous. It is. On the other hand, when the reaction temperature is too high, the polymerization rate is improved, but at the same time, thermal decomposition of the polymer during production is caused, and as a result, it becomes difficult to produce polyester having a high degree of polymerization.

  The lower limit of the reaction time is usually 2 hours or longer, and the upper limit is usually 15 hours or shorter, preferably 8 hours or shorter, more preferably 6 hours or shorter. When the reaction time is too short, a polyester with a low degree of polymerization is obtained, the tensile elongation at break is low, the terminal amount of the carboxyl group is large, and the tensile elongation at break is significantly deteriorated. There are many. On the other hand, if the reaction time is too long, the molecular weight drop due to thermal decomposition of the polyester becomes significant, and not only the tensile elongation at break is reduced, but also the amount of carboxyl group terminals that affect the durability of the polymer increases due to thermal decomposition. There is.

  In the present invention, the molar ratio of the diol component to the aliphatic dicarboxylic acid component for obtaining the polyester having the desired degree of polymerization differs in the preferred range depending on the purpose and the type of raw material, but the diol component relative to 1 mol of the acid component. The lower limit is usually 0.8 mol or more, preferably 0.9 mol or more, and the upper limit is usually 3.0 mol or less, preferably 2.7 mol or less, particularly preferably 2.5 mol or less. .

In the present invention, the polycondensation reaction is preferably performed in the presence of a polymerization catalyst. The addition timing of the polymerization catalyst is not particularly limited as long as it is before the polycondensation reaction, and it may be added when the raw materials are charged, or may be added at the start of pressure reduction.
Examples of the polymerization catalyst include compounds containing a Group 1 to Group 14 metal element excluding hydrogen and carbon in the periodic table. Specifically, at least one metal selected from the group consisting of titanium, zirconium, tin, antimony, cerium, germanium, zinc, cobalt, manganese, iron, aluminum, magnesium, calcium, strontium, sodium, and potassium. And compounds containing an organic group such as carboxylate, alkoxy salt, organic sulfonate, or β-diketonate salt, and inorganic compounds such as metal oxides and halides described above, and mixtures thereof. These catalyst components may be contained in a polyester raw material derived from biomass resources for the reasons described above. In that case, the raw material may be used as it is as a raw material containing a metal without purification. However, depending on the polyester to be produced, a polyester having a high degree of polymerization may be easier to produce as the content of a group 1 metal element such as sodium or potassium contained in the polyester raw material is smaller. In such a case, the raw material refine | purified to the grade which does not contain a group 1 metal element substantially is used suitably.

  Among these, metal compounds containing titanium, zirconium, germanium, zinc, aluminum, magnesium and calcium, and mixtures thereof are preferable, and among these, titanium compounds, zirconium compounds and germanium compounds are particularly preferable. In addition, the catalyst is preferably a compound that is liquid at the time of polymerization or that dissolves in an ester low polymer or polyester because the polymerization rate increases when it is melted or dissolved at the time of polymerization.

  The titanium compound is preferably a tetraalkyl titanate, specifically, tetra-n-propyl titanate, tetraisopropyl titanate, tetra-n-butyl titanate, tetra-t-butyl titanate, tetraphenyl titanate, tetracyclohexyl titanate, tetra Examples include benzyl titanate and mixed titanates thereof. In addition, titanium (oxy) acetylacetonate, titanium tetraacetylacetonate, titanium (diisoproxide) acetylacetonate, titanium bis (ammonium lactate) dihydroxide, titanium bis (ethylacetoacetate) diisopropoxide, titanium (triethanolaminate) ) Isopropoxide, polyhydroxytitanium stearate, titanium lactate, titanium triethanolamate, butyl titanate dimer and the like are also preferably used. Furthermore, titanium oxide and composite oxides containing titanium and silicon are also preferably used. Among these, tetra-n-propyl titanate, tetraisopropyl titanate and tetra-n-butyl titanate, titanium (oxy) acetylacetonate, titanium tetraacetylacetonate, titanium bis (ammonium lactate) dihydroxide, polyhydroxytitanium stearate Preferred are rate, titanium lactate, butyl titanate dimer, titanium oxide, titania / silica composite oxide, tetra-n-butyl titanate, titanium (oxy) acetylacetonate, titanium tetraacetylacetonate, polyhydroxy titanium stearate, titanium lactate , Butyl titanate dimer, and titania / silica composite oxide are more preferable. In particular, tetra-n-butyl titanate, polyhydroxy titanium stearate, titanium (oxy ) Acetylacetonate, titanium tetraacetyl acetonate, titania / silica composite oxide.

  Specific examples of the zirconium compound include zirconium tetraacetate, zirconium acetylate hydroxide, zirconium tris (butoxy) stearate, zirconyl diacetate, zirconium oxalate, zirconyl oxalate, potassium potassium oxalate, and polyhydroxyzirconium. Examples include stearate, zirconium ethoxide, zirconium tetra-n-propoxide, zirconium tetraisopropoxide, zirconium tetra-n-butoxide, zirconium tetra-t-butoxide, zirconium tributoxyacetylacetonate and mixtures thereof. Furthermore, zirconium oxide and composite oxides containing, for example, zirconium and silicon are also preferably used. Among these, zirconyl diacetate, zirconium tris (butoxy) stearate, zirconium tetraacetate, zirconium acetate acetate, zirconium ammonium oxalate, zirconium potassium oxalate, polyhydroxyzirconium stearate, zirconium tetra-n- Propoxide, zirconium tetraisopropoxide, zirconium tetra-n-butoxide, zirconium tetra-t-butoxide are preferred, zirconyl diacetate, zirconium tetraacetate, zirconium acetate acetate hydroxide, zirconium tris (butoxy) stearate, sulphur Zirconium ammonium acid, zirconium tetra-n-propoxide, zirconium tetra-n-butoxide are more preferred, Preferred for reasons of zirconium tris (butoxy) stearate is easily obtained polyester having a high degree of polymerization without colored.

Specific examples of the germanium compound include inorganic germanium compounds such as germanium oxide and germanium chloride, and organic germanium compounds such as tetraalkoxygermanium. In view of price and availability, germanium oxide, tetraethoxygermanium, tetrabutoxygermanium, and the like are preferable, and germanium oxide is particularly preferable.

  The amount of the catalyst used in the case of using a metal compound as these polymerization catalysts, the lower limit is usually 5 ppm or more, preferably 10 ppm or more, and the upper limit is usually 30000 ppm or less, preferably 1000 ppm or less, as the amount of metal relative to the produced polyester. More preferably, it is 250 ppm or less, Especially preferably, it is 130 ppm or less. If too much catalyst is used, it is not only economically disadvantageous but also lowers the thermal stability of the polymer. Conversely, if it is too little, the polymerization activity is lowered, and as a result, the polymer decomposes during polymer production. Is more likely to be triggered.

<Polyester and its use>
The polyester produced by the method of the present invention is usually characterized by a small amount of terminal carboxylic acid that significantly adversely affects the thermal stability of the polymer. Therefore, the polyester is excellent in thermal stability and has little deterioration in quality during molding. In the melt molding, there are few side reactions such as cleavage of terminal groups and cleavage of the main chain. Therefore, the number of terminal COOH groups of the preferred polyester produced in the present invention is usually 100 equivalent / ton (hereinafter sometimes referred to as eq / ton) or less, preferably 60 eq / ton, although it depends on the degree of polymerization of the polyester. Or less, more preferably 40 eq / ton or less, and particularly preferably 30 eq / ton or less. On the other hand, when the carboxyl group terminal amount becomes extremely small, the polymerization rate becomes extremely slow, and a polymer having a high degree of polymerization cannot be produced. For such reasons, the lower limit of the number of terminal COOH groups of the polyester is usually 0.1 eq / ton or more, more preferably 1 eq / ton.

The polyester produced in the present invention is usually preferably a polyester with little coloring. The upper limit of the yellowness (YI value) of the polyester of the present invention is usually 50 or less, preferably 30 or less, more preferably 20 or less, still more preferably 15 or less, particularly preferably 10 or less, The lower limit is not particularly limited, but is usually −20 or more, preferably −10 or more, more preferably −5 or more, particularly preferably −3 or more, and most preferably −1 or more. Polyesters exhibiting high YI values have the disadvantage that their use applications such as films and sheets are limited. On the other hand, polyesters exhibiting a low YI value are a more preferred form, but there are economical disadvantages in producing such polymers, such as complicated manufacturing processes and very high capital investment. In the present invention, the YI value is a value measured by a method based on JIS K7105. The reduced viscosity (ηsp / C) value of the polyester produced in the present invention is obtained from the reason that practically sufficient mechanical properties are obtained. 0.5 or more, preferably 1.6 or more, more preferably 2.0 or more, and particularly preferably 2.3 or more. The upper limit of the reduced viscosity (ηsp / C) value is usually 6.0 or less, preferably 5.0 or less, from the viewpoint of operability such as ease of pulling out after polyester polymerization and ease of molding. Preferably it is 4.0 or less.

The reduced viscosity as used in the present invention is measured under the following measurement conditions.
[Reduced viscosity (ηsp / C) measurement conditions]
Viscosity tube: Ubbelohde viscosity tube Measurement temperature: 30 ° C
Solvent: Phenol / tetrachloroethane (1: 1 weight ratio) solution Polyester concentration: 0.5 g / dl
To the extent that the properties of the polyester are not impaired, various additives such as a heat stabilizer, an antioxidant, a crystal nucleating agent, a difficulty are added to the reaction system in the course of the production method of the polyester of the present invention or to the obtained polyester. A flame retardant, an antistatic agent, a release agent, an ultraviolet absorber and the like may be added during polymerization.

In addition to the various additives shown above at the time of molding, a reinforcing agent such as glass fiber, carbon fiber, titanium whisker, mica, talc, CaCO ₃ , TiO ₂ , silica, and the like are added and molded. You can also.
The polyester obtained by the production method of the present invention is excellent in heat resistance and color tone, in addition, excellent in hydrolysis resistance and biodegradability, and can be produced at low cost, so it is suitable for various film applications and injection molded product applications. ing.

The molded product of the present invention can be obtained by molding the polyester of the present invention. A normal method can be adopted as the forming method. The obtained molded product is shown below together with its use.
Specific applications include injection molded products (for example, fresh food trays, fast food containers, outdoor leisure products, etc.), extruded products (films, sheets, etc., for example fishing lines, fishing nets, vegetation nets, water retaining sheets, etc.) , Hollow molded products (bottles, etc.), and other agricultural films, coating materials, fertilizer coating materials, laminate films, plates, stretched sheets, monofilaments, multifilaments, non-woven fabrics, flat yarns, staples, crimps Fiber, striped tape, split yarn, composite fiber, blow bottle, foam, shopping bag, garbage bag, compost bag, cosmetic container, detergent container, bleach container, rope, binding material, surgical thread, sanitary cover stock material It can be used for a cold box, a cushion material film, a synthetic paper, and the like.

Hereinafter, the present invention will be described in more detail with reference to examples. However, the present invention is not limited to these examples.
The quantitative analysis of acids, saccharides and cations in this example was performed using LC, and the quantitative analysis of amino acids was performed using an amino acid analyzer under the following conditions.
<Analysis of acids and sugars>
Column: ULTRON PS-80H 8.0 mmI. D. × 30cm
Eluent: Water (perchloric acid) (60% aqueous solution of perchloric acid 1.8 ml / 1 L-H ₂ O)
Temperature: 60 ° C
<Amino acid analysis>
Apparatus: Hitachi Amino Acid Analyzer L-8900
Analysis conditions: biological amino acid separation conditions-ninhydrin coloring method (570 nm, 440 nm)
Standard product: PF (Wako amino acid mixture ANII type 0.8 ml + B type 0.8 ml → 10 ml)
Injection volume: 10 μl
Quantitative calculation: Pro is calculated from the peak area of 440 nm for other amino acids and the amino acid content for other amino acids by a one-point external standard method <Cation>
Column; GL-IC-C75 (4.6 mm ID × 150 mm)
Eluent: 3.5mmol / L sulfuric acid
Column temperature: 40 ° C
On the other hand, other characteristic values were measured by the following methods.

<Nitrogen atom content>
A sample number of 10 mg was taken into a quartz boat, the sample was burned using a total nitrogen analyzer (TN-10 model manufactured by Mitsubishi Chemical Corporation), and determined by a chemiluminescence method.
<Sulfur atom content>
About 0.1 g of sample was collected in a platinum boat and burned in a quartz tube tubular furnace (AQF-100 (concentration system) manufactured by Mitsubishi Chemical Corporation), and the sulfur content in the combustion gas was 0.1% hydrogen peroxide solution. Absorbed. Then, the sulfate ion in the absorption solution was ion chromatograph (ICS manufactured by Dionex).
-1000 type).

<YI value>
It measured based on the method of JISK7105.
<Polyester reduced viscosity>
The time t (sec) during which the polyester is dissolved in phenol / tetrachloroethane (1/1 (mass ratio) mixed solution) to a concentration of 0.5 g / dL, and the solution drops in the viscosity tube in a 30 ° C. thermostat. Was measured. Further, the time t ₀ (sec) during which only the solvent falls was measured, and the reduced viscosity η _sp / C (= (t−t ₀ ) / t ₀ · C) at 30 ° C. was calculated (C is the concentration of the solution).

<Polyester terminal carboxyl group amount>
This is a value obtained by dissolving the obtained polyester in benzyl alcohol and titrating with 0.1 N NaOH, and is equivalent to a carboxyl group per 1 × 10 ⁶ g.
Reference Example <Preparation of pyruvate carboxylase (PC) enhanced strain>
(A) Extraction of genomic DNA of Brevibacterium flavum MJ233 strain Brevibacterium flavum MJ233 was established on April 28, 1975 at the Institute of Microbial Industrial Technology, Ministry of International Trade and Industry (currently National Institute of Advanced Industrial Science and Technology). Patent Biological Deposit Center) (deposited as FERM P-3068 at Tsukuba 1-1-1 Higashi 1-chome, Ibaraki, Japan 305-8586 Japan), deposited on May 1, 1981 as an international deposit under the Budapest Treaty It has been transferred and deposited with a 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 μg, thiamine 100 μg, yeast extract 1 g, casamino acid 1 g, glucose 20 g, dissolved in 1 L of distilled water] In 10 mL, Brevibacterium flavum MJ233 strain is cultured until the late logarithmic growth phase The cells were collected by centrifugation (10000 g, 5 minutes). The obtained 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. Next, proteinase K was added to the suspension so that the final concentration was 100 μg / mL, and the mixture was incubated at 37 ° C. for 1 hour. Further, 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. To this lysate, add an equal amount of phenol / chloroform solution, shake gently at room temperature for 10 minutes, and then centrifuge (5,000 × g, 20 minutes, 10-12 ° C.) to obtain a supernatant. The fraction was collected and sodium acetate was added to a concentration of 0.3 M, and then twice as much ethanol was added and mixed. The precipitate collected by centrifugation (15,000 × g, 2 minutes) was washed with 70% ethanol and then air-dried. To the obtained DNA, 5 mL of a 10 mM Tris buffer (pH 7.5) -1 mM EDTA · 2Na solution was added and left overnight at 4 ° C., and used as template DNA for subsequent PCR.

(B) Construction of plasmid for replacing PC gene promoter 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, This was performed by PCR using synthetic DNA (SEQ ID NO: 1 and SEQ ID NO: 2) designed based on the reported sequence of the gene of Corynebacterium glutamicum ATCC13032 strain (Cg0689 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 μL, PfxDNA polymerase (manufactured by Invitrogen) 0.2 μL, 1 × concentration attached buffer, 0.3 μM each primer, 1 mM MgSO ₄ , 0.25 μM dNTPs were mixed to make a total volume of 20 μL. Reaction temperature conditions:
Using 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 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. Confirmation of the amplified product was carried out by separating by 0.75% agarose (SeaKem GTG agarose: manufactured by FMC BioProducts) gel electrophoresis and then visualized by ethidium bromide staining to detect a fragment of about 0.9 kb. Recovery of the target DNA fragment from the gel was carried out using QIAQuick Gel Extraction Kit (manufactured by QIAGEN), and this was used as the PC gene N-terminal fragment.

On the other hand, a TZ4 promoter fragment that is constitutively highly expressed from Brevibacterium flavum MJ233 strain is a PCR using plasmid pMJPC1 (Japanese Patent Laid-Open No. 2005-95169) as a template and the synthetic DNAs described in SEQ ID NOs: 3 and 4 It was prepared by. The DNA of SEQ ID NO: 4 was phosphorylated at the 5 ′ end. Composition of reaction solution: 1 μL of template DNA, 0.2 μL of Pfx DNA polymerase (manufactured by Invitrogen), 1 × concentrated buffer, 0.3 μM each primer, 1 mM MgSO ₄ , 0.25 μM dNTPs were mixed to make a total volume of 20 μL. Reaction temperature condition: DNA thermal cycler PTC-200 (manufactured by MJ Research) was used, and a cycle consisting of 94 ° C. for 20 seconds, 60 ° C. for 20 seconds and 72 ° C. for 30 seconds was repeated 25 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 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, and a fragment of about 0.5 kb was detected. Recovery of the target DNA fragment from the gel was performed using QIAQuick Gel Extraction Kit (manufactured by QIAGEN), and this was used as the TZ4 promoter fragment.

  The PC gene N-terminal fragment prepared above and the TZ4 promoter fragment were mixed, and ligation kit ver. 2 (manufactured by Takara Shuzo Co., Ltd.), cleaved with restriction enzyme PstI, separated by 1.0% agarose (SeaKem GTG agarose: FMC BioProducts) gel electrophoresis, and a DNA fragment of about 1.0 kb was isolated from QIAQuick Gel Extraction Kit (Manufactured by QIAGEN) was used as a TZ4 promoter :: PC gene N-terminal fragment. Further, this DNA fragment and E. coli plasmid pHSG299 (Takara Shuzo) were mixed with DNA prepared by cutting with PstI, and ligation kit ver. 2 (Takara Shuzo) were used for connection. Escherichia coli (DH5α strain) was transformed with the obtained plasmid DNA. The recombinant Escherichia coli thus obtained was smeared on an LB agar medium containing 50 μg / mL kana machine and 50 μg / 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 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 in the 5 ′ upstream region of the pyruvate carboxylase gene derived from Brevibacterium flavum MJ233 strain was obtained using the DNA prepared in Example 1 (A) as a template and the entire genome sequence reported. It was carried out by PCR using 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: Template DNA 1 μL, PfxDNA polymerase (manufactured by Invitrogen) 0.2 μL, 1 × concentration attached buffer, 0.3 μM each primer, 1 mM MgSO ₄ , 0.25 μM dNTPs were mixed to make a total volume of 20 μL. Reaction temperature conditions: DNA thermal cycler PTC-200 (manufactured by MJ Research) was used, and a cycle consisting of 94 ° C. for 20 seconds, 60 ° C. for 20 seconds, and 72 ° C. for 30 seconds 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 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 DNA fragment of interest was recovered 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 (DH5α strain) was transformed with the resulting plasmid DNA. The recombinant Escherichia coli thus obtained was smeared on an LB agar medium containing 50 μg / mL ampicillin and 50 μg / 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 (manufactured by Takara Shuzo Co., Ltd.) 0.2 μL, 1 × concentration attached buffer, 0.2 μM each primer, 0.25 μM dNTPs were mixed to make a total volume of 20 μL. Reaction temperature conditions: DNA thermal cycler PTC-200 (manufactured by MJ Research) was used, and a cycle consisting of 94 ° C. for 20 seconds, 60 ° C. for 20 seconds and 72 ° C. for 50 seconds was repeated 20 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 5 minutes. As a result of confirming the presence or absence of the inserted DNA fragment in this way, a plasmid that recognized an amplification product of about 0.7 kb was selected and named pMJPC5.1.

  Next, the above-mentioned pMJPC17.1 and pMJPC5.1 were cleaved 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 FMC BioProducts) gel electrophoresis, and a DNA fragment of about 1.75 kb was prepared by QIAQuick Gel Extraction Kit (manufactured by QIAGEN Gel Extraction Kit). ). A DNA fragment in which the TZ4 promoter is inserted between the 5 ′ upstream region and the N-terminal region of the PC gene, and a DNA prepared by cleaving the plasmid pKMB1 (JP 2005-95169) containing the sacB gene with SacI and SphI Mixed and ligation kit ver. 2 (Takara Shuzo) were used for connection. Escherichia coli (DH5α strain) was transformed with the obtained plasmid DNA. The recombinant Escherichia coli thus obtained was smeared on an LB agar medium containing 50 μg / mL kana machine and 50 μg / 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. By cutting the obtained plasmid DNA with restriction enzymes SacI and SphI, an insert fragment of about 1.75 kb was observed, which was named pMJPC17.2 (FIG. 1).

(C) Preparation of PC-enhanced strain Plasmid DNA used for transformation of Brevibacterium flavum MJ233 / ΔLDH (lactate dehydrogenase gene-disrupted strain: Japanese Patent Laid-Open No. 2005-95169) was obtained by the calcium chloride method using plasmid DNA of pMJPC17.2. (Journal of Molecular Biology, 53, 159, 1970) was re-prepared from E. coli JM110 strain transformed. Transformation of Brevibacterium flavum MJ233 / ΔLDH strain was performed by the electric pulse method (Res. Microbiol., Vol. 144, p.181-185, 1993), and the obtained transformant contained 25 μg / 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 this medium is a plasmid in which pMJPC17.2 is a plasmid that cannot be replicated in Brevibacterium flavum MJ233 strain, so 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. Next, the homologous recombination strain was subjected to liquid culture in an LBG medium containing 25 μg / 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 removed by the second homologous recombination and sucrose-insensitive sucrose strains were obtained. Among the strains thus obtained, 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 are included. 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. . When analyzed using primers (SEQ ID NO: 8 and SEQ ID NO: 9) for PCR amplification of the TZ4 promoter and PC gene, a 678 bp DNA fragment should be observed in the promoter-substituted form. As a result of analyzing the sucrose-insensitive strain by the above method, a strain into which the TZ4 promoter was inserted was selected, and the strain was named Brevibacterium flavum MJ233 / PC-4 / ΔLDH.

(D) Measurement of pyruvate carboxylase enzyme activity The transformant Brevibacterium flavum MJ233 / PC-4 / ΔLDH strain obtained in (C) above was overnight in 100 mL of A medium containing 2% glucose and 25 mg / L kanamycin. Culture was performed. The obtained cells were collected, washed with 50 mL of 50 mM potassium phosphate buffer (pH 7.5), and resuspended in 20 mL of the same composition. The suspension was crushed with SONIFIER 350 (manufactured by BRANSON), and the centrifuged supernatant was used as a cell-free extract. The pyruvate carboxylase activity was measured using the obtained cell-free extract. The enzyme activity was measured using 100 mM Tris / HCl buffer (pH 7.5), 0.1 mg / 10 ml biotin, 5 mM magnesium chloride, 50 mM sodium bicarbonate, 5 mM sodium pyruvate, 5 mM adenosine triphosphate, 0.32 mM NADH. The reaction was carried out at 25 ° C. in a reaction solution containing 20 units / 1.5 ml malate dehydrogenase (manufactured by WAKO, derived from yeast) and the enzyme. 1 U was defined as the amount of enzyme that catalyzes the decrease of 1 μmol NADH per minute. The specific activity in the cell-free extract with enhanced pyruvate carboxylase expression was 0.1 U / mg protein. In addition, in the cells cultured in the same manner as the parent strain MJ233 / ΔLDH strain, it was below the detection limit of this activity measurement method.

Hereinafter, the Brevibacterium flavum MJ233 / PC-4 / ΔLDH strain was used as an organic acid-producing bacterium for microbial cell preparation culture and organic acid production reaction.
[Preparation of succinate-containing culture medium]
<Seed culture>
Urea: 4 g, ammonium sulfate: 14 g, monopotassium phosphate: 0.5 g, dipotassium phosphate 0.5 g, magnesium sulfate heptahydrate: 0.5 g, ferrous sulfate heptahydrate: 20 mg, sulfuric acid Manganese hydrate: 20 mg, D-biotin: 200 μg, thiamine hydrochloride: 200 μg, yeast extract: 5 g, casamino acid: 5 g, and distilled water: 100 mL of a medium of 1000 mL are placed in a 500 mL Erlenmeyer flask at 120 ° C. for 20 minutes. Heat sterilized. This was cooled to room temperature, 4 mL of a 50% aqueous glucose solution sterilized in advance was added, and Brevibacterium flavum MJ233 / PC-4 / ΔLDH constructed above was inoculated and seed-cultured at 30 ° C. for 24 hours.

<Main culture>
Ammonium sulfate: 1.0 g, monopotassium phosphate: 1.5 g, dipotassium phosphate 1.5 g, potassium chloride: 1.67 g, magnesium sulfate heptahydrate: 0.5 g, ferrous sulfate, heptahydrate Product: 40 mg, manganese sulfate / hydrate: 40 mg, D-biotin: 1.0 mg, thiamine hydrochloride: 1.0 mg, yeast extract 10 g, antifoaming agent (CE457: manufactured by NOF Corporation): 1.0 g and distilled water: 400 mL of 1000 mL of medium was placed in a 1 L fermentor and sterilized by heating at 120 ° C. for 20 minutes. After cooling to room temperature, 20 mL of 72% aqueous glucose solution sterilized in advance was added, and 20 mL of the above seed culture solution was added thereto, and the mixture was kept at 30 ° C. The pH was kept at 7.0% or less using 9.3% aqueous ammonia, main culture was performed at aeration of 300 mL / min, and stirring at 600 rpm for 24 hours. The dissolved oxygen concentration gradually decreased immediately after the start of the culture, and became almost zero 4 hours after the start of the culture. Thereafter, since the dissolved oxygen concentration increased 15 hours after the start of culture, when 380 μL of 72% glucose aqueous solution sterilized in advance was added, it rapidly decreased again and became almost zero. Since an increase in dissolved oxygen concentration was observed after about 13 minutes, 380 μL of a 72% aqueous glucose solution sterilized in advance was added and lowered again. Thereafter, a similar increase was observed about every 13 minutes, but it was reduced by the same method each time. The OD660 after 24 hours of culture was 87.3.

<Organic acid production culture>
Monoammonium phosphate: 84.4 mg, diammonium phosphate: 75.8 mg, potassium chloride 149.1 mg, magnesium sulfate heptahydrate: 0.2 g, ferrous sulfate heptahydrate: 8 mg, manganese sulfate Hydrate: 8 mg, D-biotin: 80 μg, thiamine hydrochloride: 80 μg and distilled water: 200 mL of medium was placed in a 500 mL Erlenmeyer flask and sterilized by heating at 120 ° C. for 20 minutes. After cooling to room temperature, it was placed in a 1 L jar fermenter. To 200 mL of this suspension, 90 mL of the culture solution obtained by the above main culture, 72% glucose solution sterilized in advance: 40 mL, and sterilized water: 125 mL were added and mixed, and kept at 35 ° C. The pH was maintained at 7.6 using an aqueous solution of ammonium carbonate: 154 g, 28% aqueous ammonia: 239 ml, distilled water: 650 mL, and an organic acid production reaction was performed while stirring at 200 rpm. The production succinic acid concentration at 21 hours after the start of the reaction was 34.8 g / L, and a small amount of fumaric acid was contained.

The supernatant obtained by subjecting the succinic acid fermentation broth prepared as described above to centrifugation (10,000 × G, 10 minutes) was subjected to the following Examples and Comparative Examples.
Example 1
The culture solution prepared according to the reference example was concentrated under reduced pressure heating. While stirring the concentrated culture solution, 47% sulfuric acid was added dropwise to the culture solution to adjust the pH of the solution to 2. A volume of methyl ethyl ketone (MEK) equal to that of the culture broth was added as an organic solvent to the culture broth added with sulfuric acid, and the mixture was stirred at 25 ° C. for about 30 minutes. The liquid obtained was allowed to stand, then divided into an organic layer and an aqueous layer, MEK having a volume half the volume of the aqueous layer was added to the separated aqueous layer, and the mixture was stirred at 25 ° C. for 30 minutes. Similarly, the liquid was allowed to stand, and then divided into an organic layer and an aqueous layer. The same operation was repeated three more times, and all the organic layers were combined. As a result of analyzing the organic layer by liquid chromatography (LC), 97.9% amount of succinic acid contained in the broth used was extracted from the organic layer (FIG. 2).

  Next, MEK was distilled off from the obtained MEK extract at 70 ° C. while adjusting the degree of vacuum from 400 mmHg to 100 mmHg, and the MEK extracted organic layer was concentrated. Thereafter, the obtained solution was cooled from 70 ° C. to 40 ° C. over 30 minutes, and then stirred at 40 ° C. for 1 hour. The precipitated crystals were filtered and rinsed with cold water to obtain crude crystals. On the other hand, the filtrate was further cooled from 40 ° C. to 10 ° C. over 30 minutes and stirred at 10 ° C. for 1 hour. The precipitated crystals were filtered and rinsed with cold water to obtain crude crystals. In this step, 98% of the succinic acid extracted in the MEK organic layer was recovered. Succinic acid at this stage was light brown and had a strong odor.

Next, the hydrogenation reaction of a small amount of fumaric acid contained in 150 g of crude succinic acid obtained as described above was carried out using SUS316 500 ml induction stirring autoclave in 5% Pd / C in 282 g of distilled water. (Wako catalog 326-81672, catalyst amount: 1 wt% with respect to succinic acid), hydrogen pressure was 0.2 MPa, reaction temperature was 100 ° C., and reaction time was 2 hours. As a result, all trace amounts of fumaric acid contained in the crude succinic acid (containing 0.3% by weight based on succinic acid) were all derived into succinic acid. Distilled water was added to the succinic acid solution after completion of the reaction to form a 19 wt% aqueous succinic acid solution, and then the catalyst was filtered off. The hydrogen treatment liquid had almost no odor.

  Furthermore, activated carbon (the amount of activated carbon used: 10 wt% with respect to succinic acid, Hokuetsu CL-K (activated carbon previously washed with a 5 wt% aqueous solution of succinic acid at 65 ° C) for 1 hour) was added to the solution. The activated carbon treatment of the succinic acid aqueous solution was performed under a condition of stirring at 65 ° C. for 1 hour. After the activated carbon was separated by filtration, the same treatment was performed by newly adding 10 wt% of activated carbon to the obtained succinic acid aqueous solution with respect to succinic acid. This activated carbon treatment was carried out five times in total. The absorbance at 280 nm decreased from 0.861 to 0.220 and the absorbance at 260 nm decreased from 1.258 to 0.298 after 5 times of activated carbon treatment.

The aqueous solution of succinic acid treated with activated carbon is diluted with distilled water to a concentration of 50 g / L, and a cation is contained in a trace amount by ion exchange treatment (cation exchange resin (Diaion SKT20L (Mitsubishi Chemical Corporation): H type)). Was removed.
The ion exchange treated succinic acid aqueous solution was concentrated to a succinic acid concentration of 24 wt% and crystallized. Crystallization was performed under the condition of cooling from 70 ° C. to 10 ° C. in 90 minutes and holding at 10 ° C. for 1 hour under stirring at 300 rpm using a three-one motor. The precipitated succinic acid was collected by filtration, and the crystals were washed with cold water, followed by vacuum drying at 70 ° C. for 7 hours to obtain white odorless succinic acid (YI = 2). In the obtained succinic acid, the concentrations of Na, K, Mg, Ca, and NH ₄ ions were all less than 1 ppm, the sulfur atom content was less than 1 ppm, and the nitrogen atom content was 8 ppm. In addition, a 3.0 wt% succinic acid aqueous solution of the obtained succinic acid was prepared, and the average absorbance at 250-300 nm of the spectrum measured using Hitachi spectrophotometer Hitachi UV-3500 was 0.008. (Figure 3).

[Polyester Production Example 1]
In a reaction vessel equipped with a stirrer, a nitrogen inlet, a heating device, a thermometer and a vacuum outlet, 100 parts by weight of succinic acid produced in Example 1, industrial grade 1,4-butanediol 88 manufactured by Mitsubishi Chemical Corporation .5 parts by weight, 0.37 parts by weight of malic acid and 5.4 parts by weight of 88% lactic acid aqueous solution in which 0.98% by weight of germanium dioxide was dissolved in advance as a catalyst were charged, and the system was purged with nitrogen under reduced pressure. I made it.

Next, the temperature in the system was increased to 220 ° C. while stirring, and the reaction was performed at this temperature for 1 hour. Next, the temperature was raised to 230 ° C. over 30 minutes, and at the same time, the pressure was reduced to 0.07 × 10 ³ Pa over 1.5 hours, and the reaction was performed at the same reduced pressure for 2.8 hours to complete the polymerization. As a result, white polyester (yellowness YI is 6) was obtained.
The polyester obtained had a reduced viscosity (ηsp / c) of 2.3. The terminal carboxyl group amount of the polyester was 24 equivalents / ton. The obtained polyester was uniformly dissolved in chloroform at room temperature. When chloroform was distilled off from the chloroform solution, a uniform film was formed.

Example 2
In the same manner as in Example 1, the culture solution prepared according to the reference example was concentrated under reduced pressure and heating. While stirring the concentrated culture solution, 47% sulfuric acid was added dropwise to the culture solution to adjust the pH of the culture solution to 2. A volume of methyl ethyl ketone (MEK) equal to that of the culture broth was added as an organic solvent to the culture broth added with sulfuric acid, and the mixture was stirred at 25 ° C. for about 30 minutes. The obtained liquid was allowed to stand, then divided into an organic layer and an aqueous layer, MEK having an equal volume of the aqueous layer was added to the separated aqueous layer, and the mixture was stirred at 25 ° C. for 30 minutes. Similarly, the liquid was allowed to stand, and then divided into an organic layer and an aqueous layer. The same operation was repeated once more, and all the organic layers were combined. As a result of analyzing the organic layer by liquid chromatography (LC), 97.9% amount of succinic acid contained in the broth used was extracted from the organic layer.

Next, MEK was distilled off under reduced pressure of about 50 mmHg while the organic layer was heated to 90 ° C., concentrated, cooled with ice water, and allowed to stand for 2 hours. The precipitated crystals were filtered and rinsed with cold water to obtain crude crystals. In this step, 96% of the succinic acid extracted in the MEK organic layer was recovered. Succinic acid at this stage was light brown and had a strong odor.
Next, the hydrogenation reaction of a small amount of fumaric acid contained in 150 g of crude succinic acid obtained as described above was carried out using SUS316 500 ml induction stirring autoclave in 5% Pd / C in 282 g of distilled water. (Wako catalog 326-81672, catalyst amount: 1 wt% with respect to succinic acid), hydrogen pressure was 0.2 MPa, reaction temperature was 100 ° C., and reaction time was 2 hours. As a result, all trace amounts of fumaric acid contained in the crude succinic acid (containing 0.3% by weight based on succinic acid) were all derived into succinic acid. Distilled water was added to the succinic acid solution after completion of the reaction to make a 19 wt% aqueous succinic acid solution, and then the catalyst was filtered off. The hydrogen treatment liquid had almost no odor.

Furthermore, crystallizing was performed twice. Crystallization was performed under the condition of cooling from 60 ° C. to 0 ° C. over 120 minutes and holding at 0 ° C. for 1 hour under stirring at 300 rpm using a three-one motor. The precipitated succinic acid was collected by filtration, and the crystals were washed with cold water. Further, this crystal was redissolved in distilled water to prepare a succinic acid aqueous solution having a concentration of 20 wt% succinic acid, and the same crystallization operation from 60 ° C. to 0 ° C. was performed. The obtained crystals were collected by filtration and then vacuum dried at 70 ° C. for 12 hours to obtain white odorless succinic acid (YI = 2). In the obtained succinic acid, the concentrations of Mg, Ca and NH ₄ ions are all less than 1 ppm, the sulfur atom content is less than 1 ppm, the concentrations of Na and K ions are 4 ppm and 2 ppm, respectively, and the nitrogen atom content is 9 ppm. there were. In addition, a 3.0 wt% succinic acid aqueous solution of the obtained succinic acid was prepared, and the average absorbance at 250-300 nm of the spectrum measured using Hitachi spectrophotometer Hitachi UV-3500 was 0.055. (FIG. 4).

[Polyester Production Example 2]
In a reaction vessel equipped with a stirrer, a nitrogen inlet, a heating device, a thermometer, and a vacuum outlet, 100 parts by weight of succinic acid produced in Example 2, industrial grade 1,4-butanediol 88 manufactured by Mitsubishi Chemical Corporation .5 parts by weight, 0.37 parts by weight of malic acid and 5.4 parts by weight of 88% lactic acid aqueous solution in which 0.98% by weight of germanium dioxide was dissolved in advance as a catalyst were charged, and the system was purged with nitrogen under reduced pressure. I made it.

Next, the temperature in the system was increased to 220 ° C. while stirring, and the reaction was performed at this temperature for 1 hour. Next, the temperature was raised to 230 ° C. over 30 minutes, and at the same time, the pressure was reduced to 0.07 × 10 ³ Pa over 1.5 hours, and the reaction was carried out at the same reduced pressure for 3 hours to complete the polymerization. Yellow polyester (yellowness YI is 23) was obtained.
The polyester obtained had a reduced viscosity (ηsp / c) of 2.4. The amount of terminal carboxyl groups of the polyester was 25 equivalents / ton. The obtained polyester was uniformly dissolved in chloroform at room temperature. When chloroform was distilled off from the chloroform solution, a uniform film was formed.

Example 3
The culture solution prepared according to the reference example was protonated with concentrated sulfuric acid in the same manner as in Example 1, and further extracted and crystallized to recover crude succinic acid. Succinic acid at this stage was light brown and had a strong odor.
Next, using a SUS316 500 ml induction stirring autoclave, 100 g of crude succinic acid having a strong odor obtained as described above was treated with 5% Pd / C (Wako catalog 326) in 222 g of demineralized water. -81672, catalyst amount: 1 wt% with respect to succinic acid), hydrogen pressure was 0.2 MPa, reaction temperature was 100 ° C., and reaction time was 2 hours. The hydrogen treatment liquid had almost no odor. The catalyst was separated from the treatment liquid by hot filtration at 70 ° C. The hot filtrate was cooled to 20 ° C. with stirring for about 90 minutes, and further kept at 20 ° C. for 1 hour to precipitate crystals, and solid-liquid separation was performed with Nutsche to obtain hydrous succinic acid crystals. The obtained succinic acid crystals had a white color, and no specific odor was felt from the succinic acid crystals.

Example 4
The culture solution prepared according to the reference example was protonated with concentrated sulfuric acid in the same manner as in Example 1, and further extracted and crystallized to recover crude succinic acid. Succinic acid at this stage was light brown and had a strong odor.
Next, after preparing 30 wt% crude succinic acid aqueous solution at 80 degreeC, the activated carbon Kuraray Coal GW made from Kuraray Chemical Co. of 5 wt% with respect to succinic acid was added. The activated carbon treatment was carried out for 3 hours using a constant temperature shaker kept at 80 ° C. Thereafter, the activated carbon was filtered off at 80 ° C. Even at this stage, a unique odor remained in the aqueous succinic acid solution.

  The obtained aqueous succinic acid solution was charged into a 500 ml induction stirring autoclave made of SUS316, and the hydrogen pressure was 0.8 MPa in the presence of 5% Pd / C (Wako catalog 326-81672, catalyst amount: 0.06 wt% with respect to succinic acid). The hydrogen treatment was carried out under conditions of a reaction temperature of 80 ° C. and a reaction time of 3 hours. As a result, all fumaric acid contained in the crude succinic acid in an amount of 1.8% by weight based on succinic acid was derived into succinic acid. After completion of the reaction, the catalyst was filtered off. The hydrogen treatment liquid had almost no odor.

Subsequently, the hydrogen-treated succinic acid aqueous solution was subjected to ion exchange treatment (cation exchange resin (Diaion SK1B-H (manufactured by Mitsubishi Chemical Corporation): H type)) at 80 ° C. to remove cations contained in a trace amount.
Further, the aqueous solution of succinic acid subjected to the ion exchange treatment was cooled to 20 ° C. in about 90 minutes with stirring, and further maintained at 20 ° C. for 1 hour to precipitate crystals. The precipitated succinic acid was collected by filtration, and the crystals were washed with cold water, followed by vacuum drying at 70 ° C. for 12 hours to obtain white odorless succinic acid (YI = −1). In the obtained succinic acid, the concentrations of Na, K, Mg, Ca, and NH ₄ ions were all 1 ppm or less, the sulfur atom content was less than 1 ppm, and the nitrogen atom content was 2 ppm. Moreover, the succinic acid aqueous solution of 3.0 wt% concentration of the obtained succinic acid was prepared, and the average absorbance at 250-300 nm of the spectrum measured using Hitachi UV-3500 was the succinic acid of Example 1. (0.01 or less).

[Polyester Production Example 3]
[Preparation of catalyst for polycondensation]
62.0 g of magnesium acetate tetrahydrate was placed in a 500 cm 3 glass eggplant-shaped flask equipped with a stirrer, and 250 g of absolute ethanol (purity 99% by weight or more) was further added. Further, 35.8 g of ethyl acid phosphate (mixing weight ratio of monoester and diester was 45:55) was added and stirred at 23 ° C. After confirming that magnesium acetate was completely dissolved after 15 minutes, 75.0 g of tetra-n-butyl titanate was added. Stirring was further continued for 10 minutes to obtain a uniform mixed solution. This mixed solution was transferred to a 1000 cm 3 eggplant-shaped flask and concentrated in an oil bath at 60 ° C. under reduced pressure by an evaporator. After 1 hour, most of the ethanol was distilled off, leaving a translucent viscous liquid. The temperature of the oil bath was further increased to 80 ° C., and further concentration was performed under a reduced pressure of 5 Torr. The viscous liquid gradually changed from the surface to a powder form, and was completely powdered after 2 hours. Then, it returned to normal pressure using nitrogen, and it cooled to room temperature, and obtained 108 g of pale yellow powder. The metal element analysis value of the obtained catalyst was as follows: titanium atom content was 10.3 wt%, magnesium atom content was 6.8 wt%, and phosphorus atom content was 7.8 wt%. T / P = 0.77 and M / P = 1.0. Further, a powdery catalyst was dissolved in 1,4-butanediol to prepare 34,000 ppm as titanium atoms.

[Production of aliphatic polyester resin]
A reaction vessel equipped with a stirrer, a nitrogen inlet, a heating device, a thermometer, and an exhaust port for decompression, 100 parts by weight of succinic acid produced in Example 4 as a raw material, industrial grade 1,4-butane manufactured by Mitsubishi Chemical Corporation 99.2 parts by weight of a diol and 0.38 parts by weight of malic acid (total malic acid amount 0.33 mol% with respect to succinic acid) were charged, and the system was placed in a nitrogen atmosphere by nitrogen-reduced pressure substitution.

Next, the temperature in the system was raised to 230 ° C. over 1 hour while stirring, and the reaction was carried out at this temperature for 1 hour. Thereafter, the above catalyst solution was added. The amount added was 50 ppm as titanium atoms per polyester resin obtained. The reaction temperature is gradually raised to 250 ° C., and the pressure is reduced to 0.06 × 10 ³ Pa at the same time over 2 hours. The reaction is further performed for 2.5 hours at the same reduced pressure, and the polymerization is completed. Polyester (yellowness YI is 2) was obtained.

The polyester obtained had a reduced viscosity (ηsp / c) of 2.3. The terminal carboxyl group amount of the polyester was 24 equivalents / ton.
Example 5
The culture solution prepared according to the reference example was protonated with concentrated sulfuric acid in the same manner as in Example 1, and further extracted and crystallized to recover crude succinic acid. Succinic acid at this stage was light brown and had a strong odor.

Next, a 30 wt% crude succinic acid aqueous solution was prepared at 80 ° C., and then 0.3 wt% with respect to succinic acid.
A quantity of powdered chemical activated activated charcoal diamond Hope 8ED (Mitsubishi Chemical Calgon Co., Ltd.) was added. The activated carbon treatment was performed at 80 ° C. for 2 hours while stirring at 200 rpm using a three-one motor.
After the activated carbon was filtered off at 80 ° C., the resulting aqueous succinic acid solution was charged into a 500 ml induction stirring autoclave made of SUS316, 5% Pd / C (Wako catalog 326-81672, catalyst amount: 0.06 wt% based on succinic acid) In the presence, hydrogen treatment was carried out under the conditions of a hydrogen pressure of 0.8 MPa, a reaction temperature of 80 ° C., and a reaction time of 3 hours. As a result, all fumaric acid contained in the crude succinic acid in an amount of 1.3% by weight based on succinic acid was derived into succinic acid. After completion of the reaction, the catalyst was filtered off. The hydrogen treatment liquid had almost no odor.

The hydrogenated succinic acid aqueous solution was subjected to ion exchange treatment (cation exchange resin (Diaion SK1B-H (manufactured by Mitsubishi Chemical Corporation): H type)) at 80 ° C. to remove cations contained in a trace amount.
The ion exchange treated succinic acid aqueous solution was cooled to 20 ° C. with stirring for about 90 minutes, and further kept at 20 ° C. for 1 hour to precipitate crystals. The precipitated succinic acid was collected by filtration, and the crystals were washed with cold water, followed by vacuum drying at 70 ° C. for 12 hours to obtain white odorless succinic acid (YI = −1). In the obtained succinic acid, the concentrations of Na, K, Mg, Ca, and NH ₄ ions were all 1 ppm or less, the sulfur atom content was less than 1 ppm, and the nitrogen atom content was 2 ppm. Moreover, the succinic acid aqueous solution of 3.0 wt% concentration of the obtained succinic acid was prepared, and the average absorbance at 250-300 nm of the spectrum measured using Hitachi UV-3500 was the succinic acid of Example 1. (0.01 or less).

[Polyester Production Example 4]
A polyester was produced using the same method as in Polyester Production Example 3 except that the succinic acid produced in Example 5 was used as a raw material.
In a reaction vessel equipped with a stirrer, a nitrogen inlet, a heating device, a thermometer and a vacuum outlet, 100 parts by weight of succinic acid produced in Example 5 as a raw material, 99.2 parts by weight of 1,4-butanediol, 0.38 parts by weight of malic acid (total malic acid amount 0.33 mol% with respect to succinic acid) was charged, and the system was placed in a nitrogen atmosphere by nitrogen-reduced pressure substitution.

Next, the temperature in the system was raised to 230 ° C. over 1 hour while stirring, and the reaction was carried out at this temperature for 1 hour. Thereafter, the catalyst solution described in Polyester Production Example 3 was added. The amount added was 50 ppm as titanium atoms per polyester resin obtained. The reaction temperature is gradually raised to 250 ° C., and the pressure is reduced to 0.06 × 10 ³ Pa at the same time over 2 hours. The reaction is further carried out for 2.4 hours at the same reduced pressure, and the polymerization is completed. Polyester (yellowness YI is 3) was obtained.

The polyester obtained had a reduced viscosity (ηsp / c) of 2.3. The terminal carboxyl group amount of the polyester was 24 equivalents / ton.
Comparative Example 1
The same operation as in Example 3 was performed except that the heat treatment was performed under a pressure of 0.2 MPa as the nitrogen pressure instead of performing the hydrogen treatment under a pressure of 0.2 MPa as the hydrogen pressure.

Using a 500 ml induction stirring autoclave made of SUS316, heat treatment of 100 g of crude succinic acid of the same lot as that used in Example 3 which exhibits a light brown color and exhibits a strong odor was performed in 222 g of demineralized water with 5% Pd. / C (Wako catalog 326-81672, catalyst amount: 1 wt% with respect to succinic acid), nitrogen pressure was 0.2 MPa, reaction temperature was 100 ° C., and reaction time was 2 hours. The treatment liquid had a strong odor. The catalyst was separated from the treatment liquid by hot filtration at 70 ° C. The hot filtrate was cooled to 20 ° C. with stirring for about 90 minutes, and further kept at 20 ° C. for 1 hour to precipitate crystals, and solid-liquid separation was performed with Nutsche to obtain hydrous succinic acid crystals. The obtained succinic acid crystals had a white color, but the succinic acid crystals had a strong odor peculiar to them.

Comparative Example 2
The culture solution prepared according to the reference example was protonated with concentrated sulfuric acid in the same manner as in Example 2, and further extracted and crystallized to recover crude succinic acid. Succinic acid at this stage was light brown and had a strong odor.
Next, after preparing a 20 wt% crude succinic acid aqueous solution at 80 ° C., cations contained by ion exchange treatment (cation exchange resin (Diaion SKT20L (manufactured by Mitsubishi Chemical Corporation): H type)) were removed.

Crystallization and drying were carried out from the ion exchange-treated succinic acid aqueous solution in the same procedure as in Example 2 to obtain white succinic acid (YI = 6). A characteristic odor remained in the succinic acid crystals. In the obtained succinic acid, the concentrations of Na, K, Mg, Ca, NH ₄ ions are all less than 1 ppm, the sulfur atom content is less than 1 ppm, the nitrogen atom content is 3 ppm, and the fumaric acid content is 0. 0.08% by weight. In addition, a 3.0 wt% aqueous succinic acid solution of the obtained succinic acid was prepared, and the average absorbance at 250-300 nm of the spectrum measured using Hitachi spectrophotometer Hitachi UV-3500 was 0.059. .

[Polyester production comparative example 1]
In a reaction vessel equipped with a stirrer, a nitrogen inlet, a heating device, a thermometer and a vacuum outlet, 100 parts by weight of succinic acid produced in Comparative Example 2, industrial grade 1,4-butanediol 88 manufactured by Mitsubishi Chemical Corporation .5 parts by weight, 0.37 parts by weight of malic acid and 5.4 parts by weight of 88% lactic acid aqueous solution in which 0.98% by weight of germanium dioxide was dissolved in advance as a catalyst were charged, and the system was purged with nitrogen under reduced pressure. I made it.

Next, the temperature in the system was increased to 220 ° C. while stirring, and the reaction was performed at this temperature for 1 hour. Next, the temperature was raised to 230 ° C. over 30 minutes, and at the same time, the pressure was reduced to 0.07 × 10 ³ Pa over 1.5 hours, and the reaction was performed at the same degree of pressure for 3 hours and 13 minutes to complete the polymerization. Pale yellow polyester (yellow degree YI was 34) was obtained.
The reduced viscosity (ηsp / c) of the obtained polyester was 2.2. The amount of terminal carboxyl groups of the polyester was 35 equivalents / ton. The obtained polyester was uniformly dissolved in chloroform at room temperature. When chloroform was distilled off from the chloroform solution, a uniform film was formed.

  In general, succinic acid is produced from a petrochemical-derived raw material and used in a wide variety of applications, but succinic acid derived from bioresources can be preferably used for such applications as well. . For example, 1,4-butanediol, 2-pyrrolidine, succinimide, maleic anhydride, itaconic acid, aspartic acid, maleic acid, fumaric acid, hydroxysuccinimide, maleimide, 4-aminobutyric acid, γ-aminobutyric acid, tetrahydrofuran, acrylic acid As raw materials for succinic acid esters such as dimethyl succinate and diethyl succinate, pyrrolidone, N-methylpyrrolidone, etc., as raw materials for polyesters, polyurethanes, polyamides, polymer compounds and products, acidulants, seasonings, brewing chemicals, Processed food additives, food additives, foaming bath ingredients, plant growth inhibitors, herbicides, antibacterial agents, resident mosquito attractants, etc. As raw materials and ingredients for products, etc., raw materials for products used for photography and printing, etc. Raw materials and components for metal processing, such as high-temperature welding agents and anodized surface adhesives as ingredients, and raw materials and components for adhesives and sealants, such as powdered nickel production, steel polishing baths, metal processing washing solvents, metal sintering binders, etc. As raw materials and components of solder, welding flux, porous titanium oxide production, boehmite production, photocatalyst coating agent, porous ceramic production, etc. As raw materials and components such as bleaching agents, as raw materials and components such as dyeing assistants, as raw materials and components such as electrolyte solvents and plating baths, as raw materials and components such as deodorizers and air cleaning agents, As raw materials for bioabsorbable compounds such as bioabsorbable sutures, as raw materials and ingredients for processing fiber products and softeners As raw materials and components of solvents, solvents, etc. As raw materials and components of water-soluble paint solvents, as raw materials and components of biodegradable resins, as odorless sealants, etc. As sealant raw materials and components, steel products, copper products, alloy products As a raw material and ingredients for anticorrosives, such as coating, anti-freezing, metal processing, lead for perchloric acid, boiler water treatment, synthetic lubricants, lubricants for heat-resistant plastics, lubricants for electrical contacts, etc. Ingredients and ingredients of resin, solvent for resins, polymer materials, etc., raw materials and ingredients for removal cleaning agents, etc., as raw materials and ingredients of products used in textile industry, dry cleaning, etc., solvent for ink, deinking agent , Automotive topcoat agent, insulating paint, powder paint, three-dimensional printing ink, photo-curable paint, photo-curable ink composition, nanoparticle ink, ink-jet ink, Raw materials such as oxygen-containing diesel fuel as raw materials and components for printing screen cleaning, organic semiconductor solution, ink for color filter production, toner, quinacdon pigment production, succinyl succinic acid production, dye intermediate, etc. As raw materials and components such as cement admixtures and treating agents, as raw materials and components such as engine purifiers, as raw materials and components such as petroleum refining solvents, as well as proppant resuscitation, precipitation filter cake removal, petroleum and natural As raw materials and components for gas mining aids, natural gas dehydration solvents, etc., as raw materials and components for products related to natural gas production, as low-dust concrete flooring, asphalt pavement, etc., as raw materials and components for building materials, for ink It can be used as raw materials and components such as solvents and deinking agents.

Claims (16)

A method of manufacturing a purified succinic acid from a crude succinic acid derived from biomass resources, production method of succinic acid comprising a purification step of performing hydrogen treatment in the presence of a catalyst to a solution containing at least crude succinic acid.   2. The method for producing succinic acid according to claim 1, wherein the hydrogen treatment temperature is 30 ° C. or more and 150 ° C. or less, and the hydrogen pressure is 0.1 MPa or more and 5 MPa or less.   The method for producing succinic acid according to any one of claims 1 and 2, wherein hydrogen treatment is performed with a hydrogenation catalyst in the presence of any adsorbent selected from the group consisting of metal oxide, silica and activated carbon. The method for producing succinic acid according to any one of claims 1 to 3, wherein the solution containing crude succinic acid is an aqueous solution. The method for producing succinic acid according to any one of claims 1 to 4, wherein fumaric acid is contained in a solution containing crude succinic acid to be subjected to hydrogen treatment. The method for producing succinic acid according to claim 5, wherein the content of fumaric acid in the solution containing crude succinic acid to be subjected to hydrogen treatment is 0.01 to 10% by weight based on the weight of succinic acid. As the average absorbance in the ultraviolet region of 250~300nm of succinic acid is 0.05 or less, according to claim 1, characterized in that the removal of impurities from the solution containing the crude acid A method for producing succinic acid.   The method for producing succinic acid according to claim 7, wherein the removal of the impurities is by adsorption removal using activated carbon. The method for producing succinic acid according to claim 7 , wherein the impurities are removed by crystallization using a solvent. The method for producing succinic acid according to any one of claims 1 to 9, wherein an insoluble component in the solution containing crude succinic acid is removed by membrane permeation treatment in a step prior to the hydrogen treatment step. The method for producing succinic acid according to any one of claims 1 to 9, wherein an insoluble component in the solution containing crude succinic acid is removed by an adsorbent in a step prior to the hydrogen treatment step. The crude succinic acid derived from biomass resources is subjected to hydrotreatment in the presence of a catalyst after passing through at least one of an adsorption step using activated carbon in a solution and a crystallization step. The manufacturing method of succinic acid in any one of -11.   The method for producing succinic acid according to any one of claims 1 to 12, wherein the biomass resource is a plant resource. Method for producing succinic acid according to any one of claims 1 to 13, yellowness of the purified succinic acid (YI) is equal to or is 10 or less.   The method for producing succinic acid according to any one of claims 7 to 9, wherein the impurity is a compound containing nitrogen.   The method for producing succinic acid according to any one of claims 7 to 9, wherein the impurity is an aromatic compound.