JP2021121631A - Method for producing shikimic acid - Google Patents

Abstract

To provide a method for producing shikimic acid in which shikimic acid can be obtained in good yield.SOLUTION: The method for producing shikimic acid according to the present invention comprises (E0) a step of crushing plant containing shikimic acid into a crushed product, (E1) a step of extracting shikimic acid from the crushed product, (E1a) a step of treating the shikimic acid-containing extract with plant cell wall degrading enzyme, (E2) a step of solid-liquid separating the shikimic acid-containing extract, (P1) a step of treating a solution obtained through solid-liquid separation with a hydrophobic synthetic adsorbent to yield a treated liquid, and (P2) a step of yielding a fraction containing shikimic acid from the treated liquid after performing at least the step (P1) by an ion exchange chromatography method using an amphoteric ion exchange resin, in this order.SELECTED DRAWING: None

Description

The present invention relates to a method for producing shikimic acid.

Shikimic acid, which is a natural organic compound, is an important intermediate in the biosynthesis of aromatic amino acids and alkanoids, and is industrially used as a raw material compound for the synthesis of pharmaceuticals, herbicides, antibacterial substances and the like. For example, shikimic acid is known as a raw material compound for the influenza therapeutic drug Osertamiburu.

As one of the methods for producing this shikimic acid, there is known a method of extracting shikimic acid from a plant star anise with a solvent and separating shikimic acid from an extract containing shikimic acid.
For example, Patent Document 1 discloses a method for extracting and separating star anise, which is obtained by extracting shikimic acid from a pulverized product of degreased star anise with ethanol or methanol, decolorizing it with activated carbon, and then crystallizing it from crude crystals.
Patent Document 2 discloses a method for obtaining shikimic acid by adding an ionic liquid capable of dissolving cellulose to extract shikimic acid from a plant, removing the ionic liquid with a strong basic anion exchange resin, and then obtaining shikimic acid. Has been done.

Chinese Patent No. 101168503 Japanese Unexamined Patent Publication No. 2012-188374

However, the production method described in Patent Document 1 has a problem that the yield of shikimic acid contained in a plant is low. Further, also in the production method described in Patent Document 2, the yield of shikimic acid is low, and it is necessary to use a special solvent such as an ionic liquid.
Under such circumstances, an object of the present invention is to provide a method for producing shikimic acid, which can obtain shikimic acid in good yield.

As a result of diligent studies, the present inventors have found that the following inventions meet the above object, and have arrived at the present invention.

That is, the present invention relates to the following invention.
<1> A step of recovering shikimic acid from a solution containing shikimic acid is included, and the step of recovering shikimic acid is a step of treating a solution containing shikimic acid with a hydrophobic synthetic adsorbent to obtain a treatment liquid (P1). ) And a step (P2) of obtaining a fraction containing shikimic acid from the treatment solution after performing at least the step (P1) by an ion exchange chromatography method using an amphoteric ion exchange resin. Manufacturing method.
<2> The production of shikimic acid according to <1>, which comprises a step (P3) of concentrating and drying the fraction obtained in the step (P2) after the step (P2) in the step of recovering the shikimic acid. Method.
<3> A step of obtaining a solution containing shikimic acid is included before the step of recovering the shikimic acid, and a step of obtaining the solution containing shikimic acid is a step of extracting shikimic acid from a plant containing shikimic acid. The method for producing shikimic acid according to <1> or <2>, which comprises (E1) and a step (E2) of solid-liquid separation of an extract containing shikimic acid after the step (E1).
<4> In the step of obtaining the solution containing shikimic acid, before the step (E1), a step (E0) of crushing a plant containing shikimic acid into a pulverized product is included, and the step (E1) The method for producing shikimic acid according to <3>, further comprising a step (E1a) of treating an extract containing shikimic acid with an enzyme during the step (E2).
<5> In the step of recovering the shikimic acid, the treatment solution obtained in the step (P1) is brought into contact with the strong basic anion exchange resin between the steps (P1) and the step (P2). A step (P1a) in which shikimic acid contained in the treatment liquid is adsorbed on a strong basic anion exchange resin, and then shikimic acid is eluted from the strongly basic anion exchange resin on which the shikimic acid is adsorbed with an acidic solution (P1a). The method for producing a shikimic acid according to any one of <1> to <4>, further comprising.
<6> The method for producing shikimic acid according to any one of <1> to <5>, wherein the hydrophobic synthetic adsorbent is an aromatic synthetic adsorbent.
<7> The method for producing shikimic acid according to <3> or <4>, wherein the plant containing shikimic acid is a plant of the Shikimic group group.
<8> The method for producing shikimic acid according to <4>, wherein the enzyme is a plant cell wall degrading enzyme.

<9> The steps of obtaining a solution containing shikimic acid include a step of obtaining a solution containing shikimic acid, and the steps of obtaining the solution containing shikimic acid include a step (e0) of crushing a plant containing shikimic acid into a pulverized product and the step (e0). Extracting shikimic acid from the crushed shikimic acid-containing plant obtained in the above step (e1) with a solvent containing water to obtain an extract, and treating the extract obtained in the above step (e1) with an enzyme. A method for producing shikimic acid, which comprises a step (e1a) and a step (e2) of solid-liquid separation of the extract obtained by performing the step (e1a).
<10> The step of recovering shikimic acid from the solution containing shikimic acid is included after the step of obtaining the solution containing shikimic acid, and the step of recovering shikimic acid makes the solution containing shikimic acid hydrophobic. Shikimic acid is contained in the treatment solution after at least the above step (p1) by the step (p1) of treating with a synthetic adsorbent to obtain a treatment solution and the ion exchange chromatography method using an amphoteric ion exchange resin. The method for producing shikimic acid according to <9>, which comprises a step of obtaining a fraction (p2).
<11> In the step of recovering shikimic acid from the solution containing shikimic acid, a step (p3) of concentrating and drying the fraction obtained in the step (p2) is included after the step (p2), <10>. The method for producing shikimic acid according to.
<12> In the step of recovering shikimic acid, between the step (p1) and the step (p2), the treatment solution obtained in the step (p1) is brought into contact with the strong basic anion exchange resin. A step of adsorbing shikimic acid contained in the treatment liquid on a strong basic anion exchange resin and then elution of shikimic acid from the strongly basic anion exchange resin on which the shikimic acid is adsorbed with an acidic solution (p1a). The method for producing a shikimic acid according to <10> or <11>, further comprising.
<13> The method for producing shikimic acid according to any one of <10> to <12>, wherein the hydrophobic synthetic adsorbent is an aromatic synthetic adsorbent.
<14> The method for producing shikimic acid according to any one of <9> to <13>, wherein the plant containing shikimic acid is a plant of the Shikimic group group.
<15> The method for producing shikimic acid according to any one of <9> to <14>, wherein the enzyme is a plant cell wall degrading enzyme.

<16> The method for producing shikimic acid according to <5> or <12>, wherein the strong basic anion exchange resin has a dimethylethanolammonium group.

According to the present invention, there is provided a method for producing shikimic acid, which can obtain shikimic acid in a high yield.

Hereinafter, embodiments of the present invention will be described in detail, but the description of the constituent elements described below is an example (representative example) of the embodiments of the present invention, and the present invention is described below unless the gist thereof is changed. It is not limited to the contents of.

The present invention includes a step of recovering shikimic acid from a solution containing shikimic acid, and the step of recovering shikimic acid is a step of treating the solution containing shikimic acid with a hydrophobic synthetic adsorbent to obtain a treatment liquid (a step of treating the solution containing shikimic acid with a hydrophobic synthetic adsorbent. Shikimi including P1) and a step (P2) of obtaining a fraction containing shikimic acid from the treatment solution after performing at least the above step (P1) by an ion exchange chromatography method using an amphoteric ion exchange resin. The present invention relates to a method for producing an acid (hereinafter, may be referred to as "the production method (I) of the present invention").

The present inventors have found that the amount of shikimic acid that is not recovered is large in the method of obtaining shikimic acid by treating the extract containing shikimic acid with activated carbon or crystallizing it from crude crystals, which causes a decrease in yield. I found. In the production method (I) of the present invention, by including the step (P1) and the step (P2), highly pure shikimic acid can be obtained in good yield from the solution containing shikimic acid. This is because in the production method (I) of the present invention, most of the impurities contained in the solution containing shikimic acid can be removed by treating the solution containing shikimic acid with a hydrophobic synthetic adsorbent. It is considered that this is because shikimic acid and impurities are easily separated when the shikimic acid is separated and eluted by the ion exchange chromatography method.

Further, the present invention includes a step of obtaining a solution containing shikimic acid, and the step of obtaining the solution containing shikimic acid includes a step (e0) of crushing a plant containing shikimic acid into a pulverized product and the above-mentioned step. A step (e1) of extracting shikimic acid from a plant containing the crushed shikimic acid obtained in (e0) with a solvent containing water to obtain an extract, and an enzyme of the extract obtained in the step (e1). A method for producing shikimic acid (hereinafter, "the production method (II) of the present invention"), which comprises a step (e1a) for treating with It may be described.)

The present inventors have found that when extracting from a crushed plant containing shikimic acid with a solvent containing water without using a special solvent, the filterability of the extract is low, which causes a problem. In the production method (II) of the present invention, the extraction amount of shikimic acid is improved and the filterability is improved by including the step (e0), the step (e1), the step (e1a) and the step (e2). .. This is because the fine particles (particularly fine particles of 1 μm or less) present in the extract are the cause of the decrease in the filterability of the extract obtained by extracting shikimic acid with a solvent containing water, and these fine particles are treated with an enzyme. Is thought to be due to decomposition. That is, a large amount of fine particles are present in the extract obtained by extracting shikimic acid with a solvent containing water, and these fine particles tend to clog the filter or the like during solid-liquid separation such as filtration. By treating the extract with an enzyme, the fine particles in the extract are decomposed and clogging of the filter or the like is suppressed. It is considered that this improved the filterability.

Hereinafter, the production method (I) of the present invention and the production method (II) of the present invention will be described.

[Manufacturing method (I) of the present invention]
The production method (I) of the present invention includes a step of recovering shikimic acid from a solution containing shikimic acid, and the step of recovering shikimic acid treats the solution containing shikimic acid with a hydrophobic synthetic adsorbent. A step of obtaining a fraction containing shikimic acid from the treatment solution after performing at least the above step (P1) by the step of obtaining the treatment solution (P1) and the ion exchange chromatography method using an amphoteric ion exchange resin (P1). It is a method for producing shikimic acid including P2).

[Process (P1)]
The step (P1) is a step of treating a solution containing shikimic acid with a hydrophobic synthetic adsorbent to obtain a treatment liquid.
The present inventors have found that many impurities (coloring substances, etc.) contained in a solution containing shikimic acid (particularly, an extract obtained by extracting shikimic acid from a plant containing shikimic acid) are physically contained in a hydrophobic synthetic adsorbent. It was found that it is easy to adsorb to. Therefore, by treating with a hydrophobic synthetic adsorbent, shikimic acid having a cyclohexene ring is eluted without being physically adsorbed on the hydrophobic synthetic adsorbent. On the other hand, most of the impurities can be physically adsorbed on the hydrophobic synthetic adsorbent and removed.

Examples of the solution containing shikimic acid include an extract obtained by extracting shikimic acid from a plant containing shikimic acid. The solution containing shikimic acid may be used as it is, or may be appropriately concentrated or diluted before use.
The method for preparing the solution containing shikimic acid will be described in detail later.

The "hydrophobic synthetic adsorbent" is a hydrophobic adsorbent composed of a crosslinked polymer having a porous structure, and adsorbs various organic substances in a solution by physical interaction between the pores and the substance to be adsorbed. It has the property of adsorbing.

The type of the hydrophobic synthetic adsorbent is not particularly limited as long as the object of the present invention can be achieved, and may be appropriately selected. The hydrophobic synthetic adsorbent is preferably an aromatic synthetic adsorbent in order to further improve the efficiency of removing organic impurities generated by the shikimate pathway having a benzene ring. The "aromatic synthetic adsorbent" means that the crosslinked polymer constituting the synthetic adsorbent has an aromatic ring group such as a benzene ring. Examples of the aromatic synthetic adsorbent include a porous body made of a styrene-divinylbenzene copolymer.

The pores of the hydrophobic synthetic adsorbent are not particularly limited. The hydrophobic synthetic adsorbent preferably has a pore volume of 0.5 to 3 mL / g, more preferably 1 to 3 mL / g, and even more preferably 1 to 2 mL / g. Also, the hydrophobic synthetic adsorbent is usually spherical. Further, the particle size is preferably 90% or more when it is 250 μm or more.

Specific examples of the hydrophobic synthetic adsorbent include "Diaion (registered trademark) HP20" (manufactured by Mitsubishi Chemical Corporation) and "Diaion (registered trademark) HP21" (manufactured by Mitsubishi Chemical Corporation). ..

In the present invention, the method for treating the solution containing shikimic acid with the hydrophobic synthetic adsorbent is not particularly limited as long as the solution containing shikimic acid and the hydrophobic synthetic adsorbent can be brought into contact with each other. For example, a solution containing shikimic acid in a reaction vessel, a hydrophobic synthetic adsorbent, a batch method in which a solvent is added as necessary and stirred, or a column filled with a hydrophobic synthetic adsorbent is passed through a solution containing shikimic acid. Examples thereof include a method of making a liquid.

The amount of the hydrophobic synthetic adsorbent used is not particularly limited as long as it is equal to or greater than the amount capable of adsorbing impurities contained in the solution containing shikimic acid. When the raw material is star anise, it is preferable to use 2 L or more of a hydrophobic synthetic adsorbent with respect to 1 kg of star anise.

In addition, when a solution containing shikimic acid is passed through a column packed with a hydrophobic synthetic adsorbent, an untreated liquid remains in the column when the solution containing shikimic acid is completely passed. In many cases. Therefore, it is preferable to perform cleaning using a solvent. In order to prevent almost no shikimic acid from remaining in the column, it is preferable to wash the hydrophobic synthetic adsorbent with a solvent in an amount of 1.5 volumes or more.
The solvent used for cleaning is usually water, and water such as desalted water, distilled water, and pure water can be used.

The space velocity (hereinafter referred to as SV) for flowing a solution containing shikimic acid to the hydrophobic synthetic adsorbent is usually SV = 0.5 to 5 (1 / Hr), and SV = 1 to 2 (1). / Hr) is preferable.
The SV is a unit indicating how many times the volume of the resin (synthetic adsorbent, ion exchange resin or amphoteric ion exchange resin) is passed per hour. SV is
SV = flow rate (L / Hr) / amount of filled resin (L)
Can be obtained at. For example, the space velocity (SV) when 500 mL of a resin is passed through 100 mL in 1 hour is 5 (1 / Hr).

[Process (P2)]
The step (P2) is a step of obtaining a fraction containing shikimic acid from the treatment liquid after performing at least the step (P1) by an ion exchange chromatography method using an amphoteric ion exchange resin.
By doing so, impurities not removed in the step (P1) and shikimic acid can be separated, and high-purity shikimic acid can be obtained in good yield.

In particular, in the production method (I) of the present invention, since the step (P2) is performed after the step (P1), the shikimic acid and impurities are well separated in the step (P2), and the purity of shikimic acid is 95% or more. Even when a certain fraction or a fraction having a shikimic acid purity of 98% or more is recovered, these fractions can be obtained in good yield.
Therefore, the production method (I) of the present invention is suitable as a method for producing purified shikimic acid for obtaining purified shikimic acid having a purity of 95% or more, and for producing purified shikimic acid for obtaining purified shikimic acid having a purity of 98% or more. It is more suitable as a method.
The method for measuring the purity of shikimic acid will be described in Examples.

The "amphoteric ion exchange resin" is a resin based on a crosslinked copolymer such as an acrylic or styrene resin, and has an anion exchange group and a cation exchange group.
Further, the amphoteric ion exchange resin may be a gel type or a macroporous type. Further, it may be a resin called a snake cage type, which is obtained by impregnating a strongly basic ion exchange resin with acrylic acid and polymerizing the resin.

Examples of the anion exchange group include a trimethylammonium group, a dimethylhydroxyethylammonium group, a dimethylamino group and the like. Examples of the cation exchange group include a carboxyl group, a sulfonic acid group, a phosphonic acid group, and a group forming a betaine structure combined with the above-mentioned anion exchange group.

The crosslinked copolymer, which is the resin base, is usually spherical. The average particle size (D50) is more preferably 0.05 to 2 mm, still more preferably 0.2 to 1.3 mm.

As the amphoteric ion exchange resin, an amphoteric ion exchange resin in which an ion exchange group represented by the following formula (1) is bonded to a resin base of an acrylic or styrene crosslinked copolymer is preferable.

前記式（１）において、Ｒ^１及びＲ^２は、それぞれ独立に炭素数１〜３のアルキル基である。ｋ及びｍは、それぞれ独立に１〜４の整数である。左端のアルキレン基を介して、該イオン交換基が樹脂母体に結合されている。より好ましくは、Ｒ^１及びＲ^２がメチル基であり、ｋ及びｍが１である。

In the formula (1), R ¹ and R ² are independently alkyl groups having 1 to 3 carbon atoms. k and m are independently integers of 1 to 4. The ion exchange group is bonded to the resin base via the leftmost alkylene group. More preferably, R ¹ and R ² are methyl groups, and k and m are 1.

Examples of the amphoteric ion exchange resin include "Dowex (registered trademark) Ritadion 11A8" (manufactured by Dow Chemical Co., Ltd.) and the like. Examples of the amphoteric ion exchange resin having an ion exchange group represented by the formula (1) include "Diaion (registered trademark) AMP03" (manufactured by Mitsubishi Chemical Corporation).

The treatment liquid to be applied to the step (P2) may be at least a treatment liquid after the step (P1) is performed, and even if the treatment liquid obtained in the step (P1) is used, it is added to the step (P1). You may use the treatment liquid after performing another step. Further, the treatment liquid after at least the step (P1) may be used as it is, or may be appropriately concentrated or diluted before use.

The developing solvent (eluent) is usually water, and water such as desalted water, distilled water, and pure water can be used. The amount of the developing solvent used is not particularly limited as long as it is an amount that can separate impurities and shikimic acid by an ion exchange chromatography method using an amphoteric ion exchange resin. For example, when Diaion (registered trademark) AMP03 is used as the amphoteric ion exchange resin, it is preferable to supply 1.3 times or more the volume of the developing solvent with respect to the amphoteric ion exchange resin. The space velocity (SV) for flowing the developing solvent through the amphoteric ion exchange resin is usually SV = 0.5 to 5 (1 / Hr), and SV = 1 to 2 (1 / Hr) is preferable. ..
Further, the step (P2) may be either a batch fixed layer method or a moving layer method.

Further, in the production method (I) of the present invention, it is particularly preferable to concentrate and dry the fraction obtained in the step (P2) to obtain shikimic acid. That is, it is preferable to include a step (P3) of concentrating and drying the fraction obtained in the step (P2) after the step (P2).
In the production method (I) of the present invention, by performing the step (P2) after the step (P1), a highly pure fraction of shikimic acid (for example, purified shikimic acid having a purity of 95% or more and a purity of 98%). Fractions containing the above purified shikimic acid) can be obtained. Therefore, highly pure shikimic acid can be obtained without further purification by crystallization or the like. By concentrating and drying the fraction containing shikimic acid to obtain shikimic acid, shikimic acid does not remain in the solvent without crystallization unlike the conventional purification method by crystallization, and the purity is high. Shikimic acid can be obtained in higher yield.

As the method of concentration and drying, a conventionally known method can be used. For example, methods such as vacuum heating, normal pressure heating, spray drying, drum drying, freeze drying and the like can be mentioned.

As described above, the treatment liquid to be applied to the step (P2) may be at least the treatment liquid after performing the step (P1), and the treatment liquid obtained in the step (P1) may be used as it is. , The treatment liquid after further performing a step other than the step (P1) may be used.
That is, the manufacturing method (I) of the present invention may include a step other than the step (P1) before the step (P2).

For example, a step of performing filtration using an ultrafiltration membrane may be provided between the steps (P1) and the step (P2). As the ultrafiltration membrane, one having a molecular weight cut-off of 1,000 to 100,000 or 3,000 to 80,000 can be used.

Further, in the production method (I) of the present invention, the treatment solution obtained in the step (P1) is brought into contact with the strong basic anion exchange resin between the steps (P1) and the step (P2). A step (P1a) in which shikimic acid contained in the treatment liquid is adsorbed on a strong basic anion exchange resin, and then shikimic acid is eluted from the strongly basic anion exchange resin on which the shikimic acid is adsorbed with an acidic solution (P1a). It is preferable to further contain.

[Step (P1a)]
In the step (P1a), the treatment solution obtained in the step (P1) is brought into contact with the strong basic anion exchange resin, and the shikimic acid contained in the treatment solution is adsorbed on the strong basic anion exchange resin. This is a step of elution of shikimic acid from the strongly basic anion exchange resin on which the shikimic acid is adsorbed with an acidic solution.

For example, by passing the treatment liquid obtained in the step (P1) through a column packed with a strong basic anion exchange resin, shikimic acid contained in the treatment liquid is adsorbed on the strong basic anion exchange resin. Can be made to. Further, shikimic acid can be eluted by passing an acidic solution as an eluent through the strongly basic anion exchange resin on which the shikimic acid is adsorbed.

By once adsorbing shikimic acid on the strong basic anion exchange resin and then eluting it, it is possible to separate shikimic acid from components that are not adsorbed on the strong basic anion exchange resin. The present inventors have discovered that a solution containing shikimic acid (particularly, an extract obtained by extracting shikimic acid from a plant containing shikimic acid) contains sugar as an impurity. Since this saccharide does not adsorb to the anion exchange resin, shikimic acid and the saccharide can be efficiently separated by treating with the anion exchange resin.

As described above, before the above-mentioned step (P2) (treatment with an amphoteric ion exchange resin), the step (P2) is carried out by treating with a strong basic anion exchange resin and removing impurities such as saccharides in advance. The separation of ion exchange chromatography by the amphoteric ion exchange resin in the above is better, and high-purity shikimic acid can be obtained in a higher yield. In particular, purified shikimic acid having a purity of 95% or more and purified shikimic acid having a purity of 98% or more can be obtained in a higher yield.

The "strong basic anion exchange resin" is a resin having a crosslinked polymer as a resin base and having an anion exchange group. For example, an anion exchange resin having a styrene resin as a base and a quaternary ammonium group or an amino group as an anion exchange group can be mentioned. The strong basic anion exchange resin used may be a gel type or a macroporous type, but is preferably a gel type. Further, as the strong basic anion exchange resin, a commercially available OH type resin may be used as it is, or a Cl type resin may be converted to an OH type using an aqueous solution such as sodium hydroxide and used. good.

Examples of the anion exchange group include an amino group, a trimethylammonium group, a dimethylethanolammonium group and the like. A resin in which the anion exchange group is a dimethylethanolammonium group is suitable because it can be easily regenerated (that is, Cl is converted to OH).

Specific examples of the anion exchange resin include "Diaion (registered trademark) SA20A", "Diaion (registered trademark) PA408", "Diaion (registered trademark) PA412", and "Diaion (registered trademark) PA418". (All manufactured by Mitsubishi Chemical Co., Ltd.), "Dawex (registered trademark) Marathon A2" (manufactured by Dow Chemical Co., Ltd.), "Amberlite (registered trademark) IRA410J", "Amberlite (registered trademark) IRA411", "Amberlite" (Registered Trademark) IRA910CT "(all manufactured by Organo)," Purolite (Registered Trademark) A200 "," Purolite (Registered Trademark) A300 "," Purolite (Registered Trademark) A510 "(all manufactured by Purolite)," Duo Light (registered trademark) A116 "(manufactured by Sumika Chemtex Co., Ltd.) and the like can be used.

The treatment liquid obtained in the step (P1) may be used as it is in the step (P1a), or may be appropriately concentrated or diluted. The amount of the strong basic ion exchange resin used is not particularly limited as long as it is equal to or greater than the amount that shikimic acid in the treatment liquid can be adsorbed. For example, when the raw material is octagonal, it is preferable to use 1.3 L or more of a strong basic ion exchange resin with respect to 1 kg of octagonal. The upper limit is not particularly limited, but for example, it can be 50 L or less or 20 L or less with respect to 1 kg of octagon.

The acidic solution as the eluent can be prepared using hydrochloric acid, nitric acid, sulfuric acid or the like. The concentration of the acidic solution is usually 0.5 to 3 mol / L, preferably 1 to 2 mol / L.

It is preferable to pass an eluent in an amount of 0.8 to 1.5 times the amount of the acid with respect to the total exchange capacity of the strong basic anion exchange resin. More preferably, the amount of acid is 1.05 to 1.35 times, more preferably 1.15 to 1.25 times the total exchange capacity of the strong basic anion exchange resin. .. If the amount of acid is too small, shikimic acid may not be sufficiently eluted and the yield may decrease. If the amount of the acid is too large, the separability when separating shikimic acid with the amphoteric ion exchange resin may decrease, and the yield may decrease.

Further, the space velocity (SV) is usually SV = 0.5 to 5 (1 / Hr), and SV = 1 to 2 (1 / Hr) is preferable.

The step (P1) and the step (P1a) may be carried out by using a multi-column column or the like in which columns for performing each step are connected in series.

As one of the preferred embodiments of the production method (I) of the present invention, in the step of recovering shikimic acid from the solution containing shikimic acid, the step of treating the solution containing shikimic acid with a hydrophobic synthetic adsorbent (P1). ) And the treatment solution obtained in (P1) above are brought into contact with the strongly basic anion exchange resin to adsorb the shikimic acid contained in the treatment solution to the strongly basic anion exchange resin. Obtained in the step (P1a) by elution of shikimic acid from the strongly basic anion exchange resin on which shikimic acid is adsorbed with an acidic solution and an ion exchange chromatography method using an amphoteric ion exchange resin. Examples thereof include a production method including a step (P2) of separating and elution of shikimic acid from the treated solution.

Further, the solution containing shikimic acid used in the production method (I) of the present invention is preferably a solution obtained by removing solids from an extract obtained by extracting shikimic acid from a plant containing shikimic acid. That is, the production method (I) of the present invention includes a step of obtaining a solution containing shikimic acid before the step of recovering shikimic acid from the solution containing shikimic acid, and prepares the solution containing shikimic acid. The obtaining step is a production method including a step of extracting shikimic acid from a plant containing shikimic acid (E1) and a step of solid-liquid separating the extract containing shikimic acid after the step (E1) (E2). be able to.

[Process (E1)]
Step (E1) is a step of extracting shikimic acid from a plant containing shikimic acid. Specifically, it is a step of mixing a plant containing shikimic acid and a solvent and extracting shikimic acid from the plant containing shikimic acid into the solvent.

Examples of the plant containing shikimic acid include a plant belonging to the Ginkgo family, a plant belonging to the Shikimi group, and the like, and a plant belonging to the Shikimi group is preferable.
Examples of plants belonging to the Illicium family include Illicium floridanum, Illicium diffengri, Illicium henryi, Illicium verum, Illicium lancealatum, Illicium illicium, and Illicium.

From the viewpoint of the content of shikimic acid and the presence or absence of toxic components, the plant containing shikimic acid is preferably selected from the group consisting of Illicium henryi, Illicium verum and Illicium pachyphyllum, and is more preferably the fruit portion of these plants. preferable.

One of the particularly suitable raw materials is dried fruit of Illicium verum called star anise.

Further, in the step (E1), the shape of the plant containing shikimic acid as a raw material is not particularly limited. The fruit portion of a plant belonging to the genus Illicium may be used as it is, or it may be cut or crushed to an appropriate size before use.

In order to further improve the extraction efficiency, the shape of the plant is preferably powdery. That is, in the step of obtaining the solution containing shikimic acid, it is preferable to further include the step (E0) of crushing the plant containing shikimic acid into a pulverized product before the step (E1).
The crushing method is not particularly limited as long as it can crush a plant containing shikimic acid, and a conventionally known method can be used. For example, a crushing device using a compressive force, a shearing force, an impact force, a frictional force, or the like can be used. Specific examples thereof include a jaw crusher, a roll crusher, a screw mill, and a ball mill.

Water, alcohol, or the like can be used as the extraction solvent. From the viewpoint of ease of removal of oil components contained in plants containing shikimic acid and safety, it is preferable that the solvent contains water as a main component (50% by mass or more), and water is particularly preferable.

The amount of extraction solvent is appropriately determined. If the amount of the extraction solvent is too small, the extraction efficiency may decrease. Therefore, the amount of the extraction solvent is preferably 5 times by mass or more, more preferably 7 times by mass or more with respect to the plant containing shikimic acid. On the other hand, even if the amount of the extraction solvent is too large, it takes time for solid-liquid separation and the like. In addition, it becomes a factor of cost increase. Therefore, the upper limit of the amount of the extraction solvent is preferably 15 times by mass or less, and more preferably 10 times by mass or less with respect to the plant containing shikimic acid.

The extraction temperature is appropriately determined according to the extraction solvent and the like within a range in which shikimic acid can be extracted, and may be 90 ° C. or higher, but is preferably 100 ° C. or higher. If the extraction temperature is too low, the extraction efficiency may decrease. On the other hand, the upper limit of the extraction temperature is arbitrary as long as it is below the temperature at which shikimic acid does not decompose or deteriorate. For example, the temperature can be 200 ° C. or lower.

The extraction time is appropriately determined according to the amount of the raw material and the extraction solvent. In addition, plants containing shikimic acid usually contain an oil component, and if this oil component remains in the extract, solid-liquid separation may become difficult due to deterioration of filterability and the like. Therefore, it is preferable to extract shikimic acid from the plant containing shikimic acid and at the same time remove the oil component contained in the plant containing shikimic acid. The extraction time is preferably longer than the time during which shikimic acid can be extracted and the oil component contained in the plant containing shikimic acid can be removed. For example, the extraction time may be 1 hour or more, 5 hours or more, or 10 hours or more. The upper limit is also not particularly limited, but may be, for example, 48 hours or less or 24 hours or less in consideration of side reactions and the like.

For extraction of shikimic acid and removal of oil components, for example, when the extraction solvent is water, the extraction solvent and the oil component are co-boiled in the tank in which the extraction operation is performed, and the extraction solvent and the oil component are distilled out of the tank. As an example, a method of returning the extraction solvent of the lower layer portion of the distilled distillate to the inside of the tank can be given. By doing so, it is possible to extract shikimic acid and remove oil components while keeping the liquid amount of the extraction solvent constant.

[Process (E2)]
The step (E2) is a step of solid-liquid separating the extract containing shikimic acid, and is a step of removing solids such as crushed plants containing shikimic acid from the extract containing shikimic acid.
The method of solid-liquid separation is not particularly limited, and a conventionally known method can be used. For example, solid-liquid separation can be performed by filtration using an arbitrary filter medium, vacuum filtration, centrifugation or the like. Preferably, it is filtration using an arbitrary filter medium.

Further, in the production method (I) of the present invention, it is preferable to further include a step (E1a) of treating the extract containing shikimic acid with an enzyme before the step (E2). The enzyme treatment can be performed before the step (E1), but is preferably performed between the steps (E1) and the step (E2). By doing so, the filterability of the extract during solid-liquid separation can be improved.

The enzyme is preferably a plant cell wall degrading enzyme.
Examples of the enzyme that decomposes the cell wall of a plant include cellulase, pectinase, xylanase, lysozyme, and the like, and preferably contains cellulase and / or pectinase.
In addition, two or more types of enzymes may be used in combination. When two or more kinds of enzymes are used, the enzyme treatment may be carried out by adding two or more kinds of enzymes at the same time, or the enzyme treatment may be carried out a plurality of times by using each enzyme individually.

The enzyme treatment is usually carried out by dissolving or dispersing the enzyme in the extract and stirring for an arbitrary time. The enzyme treatment temperature is appropriately determined according to the type of enzyme used, the shape of the raw material, and the like within a range that does not impair the object of the present invention, but is usually 35 ° C to 60 ° C.
The enzyme treatment time is appropriately determined depending on the enzyme treatment temperature and the like, and is preferably 0.5 hours or more. The upper limit of the enzyme treatment time is not particularly limited, but may be, for example, 24 hours or less or 15 hours or less.

The amount of the enzyme added is appropriately determined according to the enzyme to be used and the like, and is preferably 0.05% by mass or more with respect to 100% by mass of the plant to be extracted (plant containing shikimic acid) in the step (E1). .. The amount of the enzyme added is not particularly limited, but if it exceeds a certain amount, the effect may be diminished. Therefore, the amount of the enzyme added is 10% by mass or less with respect to 100% by mass of the plant to be extracted (plant containing shikimic acid). It is preferably 5% by mass or less, and more preferably 5% by mass or less.

Further, it is preferable that the enzyme treatment is followed by further heat treatment. The temperature at the time of this heat treatment is not particularly limited as long as it is equal to or higher than the temperature at which the enzyme can be inactivated. For example, it can be 60 ° C. or higher or 90 ° C. or higher. The upper limit of the heat treatment temperature may be a temperature below which shikimic acid does not decompose or denature. For example, it may be 200 ° C. or lower or 150 ° C. or lower.
Further, it is sufficient that the enzyme can be inactivated during the heat treatment time. For example, it is preferably 20 minutes or more, and more preferably 30 minutes or more. If the heating time is too short, the enzyme may not be sufficiently inactivated. The upper limit of the heating time is not particularly limited, and may be 24 hours or less, 12 hours or less, or 5 hours or less.
By performing the heat treatment, the yield of shikimic acid in the step (P1) and the step (P2) is further improved.

In particular, in the production method (I) of the present invention, the solution containing shikimic acid is prepared by adding a plant containing shikimic acid before the step (E1) in addition to the step (E1) and the step (E2). Obtained by a method including a step (E0) of pulverizing to obtain a pulverized product, and further including a step (E1a) of treating an extract containing shikimic acid with an enzyme between the steps (E1) and the step (E2). Is preferable. That is, a step (E0) of crushing a plant containing shikimic acid into a crushed product, a step of extracting shikimic acid from the crushed product (E1'), and an extract obtained in the step (E1') are used. It is preferable to use a solution obtained by a method including a step of treating with an enzyme (E1a') and a step of solid-liquid separation of the extract obtained by performing the step (E1a') (E2).

The step of obtaining the solution containing shikimic acid and the step of recovering shikimic acid from the solution containing shikimic acid may be continuously performed or may be performed after a predetermined time.

[Manufacturing method (II) of the present invention]
The production method (II) of the present invention includes a step of obtaining a solution containing shikimic acid, and the step of obtaining the solution containing shikimic acid is a step of crushing a plant containing shikimic acid into a pulverized product (e0). And the step (e1) of extracting shikimic acid from the crushed shikimic acid-containing plant obtained in the step (e0) with a solvent containing water to obtain an extract, and the step (e1) obtained in the step (e1). This is a method for producing shikimic acid, which comprises a step of treating the extract with an enzyme (e1a) and a step of solid-liquid separation of the extract obtained by performing the step (e1a) (e2).

[Step e0]
The step (e0) is a step of crushing a plant containing shikimic acid into a crushed product. Extraction efficiency can be improved by crushing plants containing shikimic acid into powder.

The plant containing shikimic acid, which is a raw material for obtaining a plant containing shikimic acid, is the same as the production method (I) of the present invention, and is an octagon (or star anise) as in the production method (I) of the present invention. A dried fruit of Illicium verum called is one of the suitable raw materials.

The method for crushing a plant containing shikimic acid is not particularly limited as long as it can crush a plant containing shikimic acid, and a conventionally known method can be used. For example, a crushing device using a compressive force, a shearing force, an impact force, a frictional force, or the like can be used. Specific examples thereof include a jaw crusher, a roll crusher, a screw mill, and a ball mill.

[Step (e1)]
The step (e1) is a step of extracting shikimic acid from the crushed shikimic acid-containing plant obtained in the step (e0) with a solvent containing water. Specifically, it is a step of mixing a plant containing shikimic acid and a solvent and extracting shikimic acid from the plant containing shikimic acid into the solvent.

The extraction solvent may be water alone, or may contain other solvents as long as water is the main component (50% by mass or more). From the viewpoint of ease of removal and safety of oil components contained in plants containing shikimic acid, water is preferable.

The amount of extraction solvent is appropriately determined. If the amount of the extraction solvent is too small, the extraction efficiency may decrease. Therefore, the amount of the extraction solvent is preferably 5 times by mass or more, more preferably 7 times by mass or more with respect to the plant containing shikimic acid. On the other hand, even if the amount of the extraction solvent is too large, it takes time for solid-liquid separation and the like. In addition, it becomes a factor of cost increase. Therefore, the upper limit of the amount of the extraction solvent is preferably 15 times by mass or less, and more preferably 10 times by mass or less with respect to the plant containing shikimic acid.

The extraction temperature is appropriately determined according to the extraction solvent and the like within a range in which shikimic acid can be extracted, and may be 90 ° C. or higher, but is preferably 100 ° C. or higher. If the extraction temperature is too low, the extraction efficiency may decrease. On the other hand, the upper limit of the extraction temperature is arbitrary as long as it is at least a temperature at which shikimic acid does not decompose or deteriorate, and can be, for example, 200 ° C. or less.

The extraction time is appropriately determined according to the amount of the raw material and the extraction solvent. In addition, plants containing shikimic acid usually contain an oil component, and if this oil component remains in the extract, solid-liquid separation may become difficult due to deterioration of filterability and the like. Therefore, it is preferable to extract shikimic acid from the plant containing shikimic acid and at the same time remove the oil component contained in the plant containing shikimic acid. The extraction time is preferably longer than the time during which shikimic acid can be extracted and the oil component contained in the plant containing shikimic acid can be removed. For example, the extraction time may be 1 hour or more, 5 hours or more, or 10 hours or more. The upper limit is not particularly limited, but may be 48 hours or less or 24 hours or less in consideration of side reactions and the like.

For the extraction of shikimic acid and the removal of the oil component, for example, the extraction solvent and the oil component are azeotroped in the tank in which the extraction operation is performed, and the extraction solvent and the oil component are distilled out of the tank, and the distilled distillate is distilled. A method of returning the extraction solvent in the lower layer of the effluent to the inside of the tank can be mentioned. By doing so, it is possible to extract shikimic acid and remove oil components while keeping the liquid amount of the extraction solvent constant.

[Step (e1a)]
The step (e1a) is a step of treating the extract obtained in the step (e1) with an enzyme.
The enzyme is preferably a plant cell wall degrading enzyme.
Examples of the enzyme that decomposes the cell wall of a plant include cellulase, pectinase, xylanase, lysozyme, and the like, and preferably contains cellulase and / or pectinase.
In addition, two or more types of enzymes may be used in combination. When two or more kinds of enzymes are used, the enzyme treatment may be carried out by adding two or more kinds of enzymes at the same time, or the enzyme treatment may be carried out a plurality of times by using each enzyme individually.

The enzyme treatment is usually carried out by dissolving or dispersing the enzyme in the extract and stirring for an arbitrary time. The enzyme treatment temperature is appropriately determined according to the type of enzyme used, the shape of the raw material, and the like within a range that does not impair the object of the present invention, but is usually 35 ° C to 60 ° C.
The enzyme treatment time is appropriately determined depending on the enzyme treatment temperature and the like, and is preferably 0.5 hours or more. The upper limit of the enzyme treatment time is not particularly limited, but may be, for example, 24 hours or less or 15 hours or less.

The amount of the enzyme added is appropriately determined according to the enzyme to be used and the like, and is preferably 0.05% by mass or more with respect to 100% by mass of the plant to be extracted (plant containing shikimic acid) in the step (e1). .. The amount of the enzyme added is not particularly limited, but if it exceeds a certain amount, the effect may be diminished. Therefore, the amount of the enzyme added is 10% by mass or less with respect to 100% by mass of the plant to be extracted (plant containing shikimic acid). It is preferably 5% by mass or less, and more preferably 5% by mass or less.

Further, it is preferable that the enzyme treatment is followed by further heat treatment. The temperature at the time of this heat treatment is not particularly limited as long as it is equal to or higher than the temperature at which the enzyme can be inactivated. For example, it can be 60 ° C. or higher or 90 ° C. or higher. The upper limit of the heat treatment temperature may be a temperature below which shikimic acid does not decompose or denature. For example, it may be 200 ° C. or lower or 150 ° C. or lower.
Further, it is sufficient that the enzyme can be inactivated during the heat treatment time. For example, it is preferably 20 minutes or more, and more preferably 30 minutes or more. If the heating time is too short, the enzyme may not be sufficiently inactivated. The upper limit of the heating time is not particularly limited, and may be 24 hours or less, 12 hours or less, or 5 hours or less.

[Step (e2)]
The step (e2) is a step of solid-liquid separating the extract obtained by performing the step (e1a), and is a step of removing solid content such as crushed plants containing shikimic acid from the extract.
The method of solid-liquid separation is not particularly limited, and a conventionally known method can be used. For example, solid-liquid separation can be performed by filtration using an arbitrary filter medium, vacuum filtration, centrifugation or the like. Preferably, it is filtration using an arbitrary filter medium.

Further, the production method (II) of the present invention usually includes a step of recovering shikimic acid from the solution containing shikimic acid after the step of obtaining a solution containing shikimic acid, and the step of recovering the shikimic acid is included. After performing at least the above steps (p1) by a step (p1) of treating a solution containing shikimic acid with a hydrophobic synthetic adsorbent to obtain a treatment solution and an ion exchange chromatography method using an amphoteric ion exchange resin. It is preferable to include a step (p2) of obtaining a fraction containing shikimic acid from the treatment solution of.

Further, in the step of recovering shikimic acid from the solution containing shikimic acid, it is preferable to include a step (p3) of concentrating and drying the fraction obtained in the step (p2) after the step (p2).

The step (p1) is the same as the step (P1) in the manufacturing method (I) of the present invention. The step (p2) is the same as the step (P2) in the manufacturing method (I) of the present invention. The step (p3) is the same as the step (P3) in the manufacturing method (I) of the present invention.

Further, a step other than the step (p1) may be included before the step (p2). For example, a step of performing filtration using an ultrafiltration membrane may be provided between the steps (p1) and the step (p2). As the ultrafiltration membrane, one having a molecular weight cut-off of 1,000 to 10,000 or 3,000 to 80,000 can be used.

Further, in the production method (II) of the present invention, the step of recovering the shikimic acid from the solution containing the shikimic acid is obtained in the step (p1) between the step (p1) and the step (p2). The treatment solution was brought into contact with the strong basic anion exchange resin, and the shikimic acid contained in the treatment solution was adsorbed on the strong basic anion exchange resin, and then the strong basic anion exchange to which the shikimic acid was adsorbed was adsorbed. It is preferable to further include a step (p1a) of elution of shikimic acid from the resin with an acidic solution. The step (p1a) is the same as the step (P1a) in the manufacturing method (I) of the present invention.

Hereinafter, the present invention will be described in more detail with reference to Examples, but the present invention is not limited to the following Examples unless the gist thereof is changed.

[Example 1]
(Step to obtain a solution containing shikimic acid)
In a 2 L 4-neck flask equipped with a stirrer, a condenser, and a Dean-Stark trap, 150 g of star anise and 1050 g of demineralized water were placed, and the mixture was refluxed at 100 ° C. for 6 hours to extract shikimic acid contained in the star anise. 1a) was obtained. During reflux, the oil component and water (distillate water) contained in the star anise accumulated in the Dean-Stark trap. The reflux was carried out while appropriately extracting the distillate water and returning the distillate water from another mouth of the flask into the flask so that the oil component accumulated in the Dean-Stark trap did not return to the flask. In addition, the oil component accumulated in the Dean-Stark trap was removed.
After cooling to 40 ° C., an enzyme (0.75 g of sucrase C manufactured by Mitsubishi Chemical Foods, 0.75 g of sucrase N) was added to the extract (1a), and the mixture was stirred at 40 ° C. for 12 hours. Then, the mixture was stirred at 90 ° C. for 30 minutes to inactivate the enzyme to obtain an extract (1b).
After cooling to 30 ° C., the extract (1b) is filtered (filter paper, Advantech No5C, 110 mm) and washed with 200 mL of desalted water to remove solid components contained in the extract (1b) and extract containing shikimic acid. Liquid (1) 1500 mL was obtained.
As the star anise, Chinese star anise (star anise (hall) manufactured by S & B Foods Co., Ltd.) was used as it was.

(Step of recovering shikimic acid from a solution containing shikimic acid)
A column (1) having an inner diameter of 4.5 cm was filled with 300 mL (inner diameter 4.5 x height 23 cm) of a hydrophobic synthetic adsorbent (manufactured by Mitsubishi Chemical Corporation, Diaion (registered trademark) HP20). 1500 mL of the obtained extract (1) was passed through the column (1) at a flow rate of SV = 1 with a roller pump, and then desalted water was passed through the column (1) to obtain 1700 mL of the treatment liquid (1).

Next, the treatment liquid (1) was treated with an ultrafiltration membrane using a UF membrane module (Asahi Kasei Chemicals, Inc., Microza (registered trademark) UF pencil type module SLP-0053, molecular weight cut-off 10,000). , 1650 mL of permeate (1) was obtained. The permeate (1) was concentrated to 110 mL using an evaporator to obtain a concentrate (1). This concentrated solution (1) was used as a stock solution for treatment with an amphoteric ion exchange resin.

A column (3) having an inner diameter of 5 cm was filled with 500 mL (inner diameter 5 cm × 31.5 cm) of an amphoteric ion exchange resin (Diaion (registered trademark) AMP03, manufactured by Mitsubishi Chemical Corporation). 110 mL of the concentrated solution (1) was passed through the column (3), and further, 1600 mL of demineralized water was passed through the column (3). The concentrated solution (1) and desalinated water were supplied by a roller pump at a flow rate of SV = 1 (1 / Hr).
Of the liquids eluted from the column (3), the first 600 mL of the eluate was discarded and the next 1000 mL of the eluate (1) was collected. This eluate (1) was concentrated with an evaporator and then dried in a vacuum dryer (full vacuum, 50 ° C.) for 13 hours to obtain 4.8 g of shikimic acid having a purity of 98% or more.

[Example 2]
(Step to obtain a solution containing shikimic acid)
200 g of star anise was put into a carita coffee mill (classic mill) and crushed to obtain a powdered star anise.
150 g of crushed star anise powder and 1050 g of demineralized water were placed in a 2 L four-necked flask equipped with a stirrer, a condenser, and a Dean-Stark trap, and the mixture was refluxed at 100 ° C. for 6 hours to extract shikimic acid contained in the star anise. An extract (2a) was obtained. During reflux, the oil component and water (distillate water) contained in the star anise accumulated in the Dean-Stark trap. The reflux was carried out while appropriately extracting the distillate water and returning the distillate water from another mouth of the flask into the flask so that the oil component accumulated in the Dean-Stark trap did not return to the flask. In addition, the oil component accumulated in the Dean-Stark trap was removed.
After cooling to 40 ° C., enzymes (0.75 g of sucrase C and 0.75 g of sucrase N manufactured by Mitsubishi Chemical Foods) were added to the extract (2a), and the mixture was stirred at 40 ° C. for 12 hours. Then, the mixture was stirred at 90 ° C. for 30 minutes to inactivate the enzyme to obtain an extract (2b).
After cooling to 30 ° C., the extract (2b) is filtered (filter paper, Advantech No5C, 110 mm), washed with 200 mL of desalted water to remove solid components contained in the extract (2b), and the extract (2). 1500 mL was obtained.

(Step of recovering shikimic acid from a solution containing shikimic acid)
Purification was carried out by the same method as in the step of recovering shikimic acid from the solution containing shikimic acid in Example 1 except that the extract (2) was used instead of the extract (1), and the purity was 98% or more. 6.2 g of shikimic acid was obtained.

[Example 3]
(Step to obtain a solution containing shikimic acid)
An extract of shikimic acid (3) was obtained in the same manner as in the step of obtaining a solution containing shikimic acid in Example 2.

(Step of recovering shikimic acid from a solution containing shikimic acid)
A column (1) having an inner diameter of 4.5 cm was filled with 300 mL (inner diameter 4.5 x height 23 cm) of a hydrophobic synthetic adsorbent (manufactured by Mitsubishi Chemical Corporation, Diaion (registered trademark) HP20). Further, a column (2) having an inner diameter of 4.5 cm is filled with 200 mL of a Cl-type anion exchange resin (Diaion (registered trademark) SA20A, manufactured by Mitsubishi Chemical Co., Ltd.) and treated with an aqueous sodium hydroxide solution (35 g / 500 ml). Then, it was made into an OH type anion exchange resin. Next, the column (1) and the column (2) were connected in series so that the treatment liquid treated by the column (1) could be treated by the column (2), and a two-bed, two-tower type device was assembled. ..
First, 1500 mL of the obtained extract containing shikimic acid (3) was passed through a two-bed, two-tower device. At this time, shikimic acid passed through the hydrophobic synthetic adsorbent of the column (1) without being adsorbed, was supplied to the column (2), and was adsorbed on the SA20A.
Next, 230 mL of 1N hydrochloric acid was passed through the column (2) of SA20A on which shikimic acid was adsorbed, and 865 mL of desalinated water was further passed to obtain 1100 mL of eluent. The liquid was supplied by a roller pump at a flow rate of SV = 1 (1 / Hr). The obtained eluate (3) was concentrated with an evaporator to obtain 110 mL of the concentrate (3).

110 mL of the concentrated solution (3) was passed through a column (3) filled with 500 mL (inner diameter 5 cm × 31.5 cm) of an amphoteric ion exchange resin (Diaion (registered trademark) AMP03, manufactured by Mitsubishi Chemical Corporation), and further. 1600 mL of demineralized water was passed through. Of the eluates, the first 600 mL of eluate was discarded and the next 1000 mL of eluate (3) was collected. This eluate (3) was concentrated with an evaporator and then dried in a vacuum dryer (full vacuum, 50 ° C.) for 13 hours to obtain 12.8 g of shikimic acid having a purity of 98% or more.

[Example 4]
(Step to obtain a solution containing shikimic acid)
The coffee mill was adjusted so as to be finely crushed as compared with Example 2, and the shikimic acid was contained in the same manner as in the step of obtaining the solution containing shikimic acid except that the plant containing shikimic acid was crushed. The extract (4) was obtained.

(Step of recovering shikimic acid from a solution containing shikimic acid)
Purification was carried out in the same manner as in the step of recovering shikimic acid from the solution containing shikimic acid in Example 3 except that the extract (4) was used instead of the extract (3). 14.3 g of acid was obtained.

[Comparative Example 1]
200 g of star anise was put into a carita coffee mill and crushed to obtain a powdery star anise.
362 g of crushed star anise powder and 1009 g of methanol (manufactured by Nippon Steel Chemical Co., Ltd.) were placed in a 2 L four-necked flask equipped with a stirrer and a condenser, and refluxed at 65 ° C. for 30 minutes to obtain an extract (5a).
After cooling to 30 ° C., the extract (5a) was filtered (filter paper, Advantec No. 5C, 110 mm) to obtain 653 g of the extract (5b).
This extract (5b) and 171 g of petroleum ether (manufactured by JXTG Energy Co., Ltd., reagent ligroin special grade) are placed in a separating funnel, shaken well for about 5 minutes, and then the lower phase (methanol phase) of the separated phase is extracted (5c). ) Was collected.
After mixing 322 g of demineralized water with the recovered extract (5c), filtration (filter paper, Advantec No. 5C, 110 mm) was performed to obtain 375 g of the extract (5d).

Further, 21 g of activated carbon (Carborafine, manufactured by Japan Enviro Chemicals, Inc.) was mixed with this extract (5d), and the mixture was stirred at 60 ° C. for 20 minutes.
After cooling to 30 ° C., the extract (5d) after the activated carbon treatment was filtered (filter paper, Advantec No. 5C, 110 mm) to obtain 319 g of the extract (5e).
The extract (5e) was concentrated at 60 ° C. and 80 Torr to obtain 25 g of the concentrated solution (5).
A small amount of shikimic acid was added as a seed crystal to this concentrated solution (5), and the mixture was left in a cool and dark place for 24 hours. Then, it was filtered (filter paper, Advantech No5C, 110 mm) and washed with ethanol water (20 g of ethanol (manufactured by Japan Alcohol Trading Co., Ltd.) + 3 g of ultrapure water). Further, the mixture was washed with ethanol water (15 g of ethanol + 3 g of ultrapure water) to obtain 62 g of an extract (5f).
The wet shikimic acid on the filter paper was dried (full vacuum in a vacuum dryer, treated at 60 ° C. for 13 hours) to obtain 3.6 g of shikimic acid having a purity of 93.8%.

[Comparative Example 2]
200 g of star anise was put into a carita coffee mill (classic mill) and crushed to obtain a powdered star anise.
150 g of crushed star anise powder and 1050 g of demineralized water were placed in a 2 L round-bottom flask and refluxed at 100 ° C. for 6 hours to obtain an extract (6a).
After cooling to 30 ° C., the extract (6a) was filtered (filter paper, Advantec No5C, 110 mm), but the filter paper was clogged with fine particles and could not be filtered.

The purity, extraction rate and filterability of shikimic acid of Examples 1 to 4 and Comparative Example 1 were evaluated.
[purity]
The shikimic acid concentration was determined by the HPLC method (column: Inertsil ODS-3 (particle size 3 μm, length 150 mm, inner diameter 4.6 mm), mobile phase: (A) CH ₃ CN + (B) 2.1% acetic acid solution (A /). B = 30/70 (v / v)), 0.5 mL / min, column temperature: 30 ° C., detection: 254 nm).
For the measurement, a calibration curve was prepared using a shikimic acid solution (solvent: ultrapure water) of 100 to 1000 ppm.
The shikimic acid content was calculated from the shikimic acid concentration obtained by the measurement, and the purity of shikimic acid was determined by the following formula.
Purity of shikimic acid = content of shikimic acid / sample weight x 100 (%)

[Extraction rate]
The extraction rate was calculated as follows. The results are shown in Table 1.
The extraction rate was calculated by the following formula.
Extraction rate = weight of extracted shikimic acid / weight of raw material (%)

[Filtability]
The filterability at the time of removing the solid component contained in the extract in the step of obtaining the solution containing shikimic acid of Examples 1 to 4 and Comparative Examples 1 and 2 was evaluated according to the following criteria.
The results are shown in Table 1.
×: Time required for filtration of 1500 mL is 2 days or more Δ: Time required for filtration of 1500 mL is within 3 hours 〇: Time required for filtration of 1500 mL is within 1 hour

According to the present invention, shikimic acid can be obtained in high yield, and the obtained shikimic acid can be used as a synthetic raw material for various substances, which is industrially useful.

Claims (6)

The step (E0) of crushing a plant containing shikimic acid into a crushed product,
The step (E1) of extracting shikimic acid from the pulverized product and
The step of treating the extract containing shikimic acid with a plant cell wall degrading enzyme (E1a), and
The step (E2) of solid-liquid separation of the extract containing shikimic acid and
The step (P1) of treating the solution obtained through the solid-liquid separation with a hydrophobic synthetic adsorbent to obtain a treated liquid, and
A step (P2) of obtaining a fraction containing shikimic acid from the treatment liquid after performing at least the above step (P1) by an ion exchange chromatography method using an amphoteric ion exchange resin.
A method for producing shikimic acid, which comprises in this order. In the step of recovering shikimic acid,
The method for producing shikimic acid according to claim 1, further comprising a step (P3) of concentrating and drying the fraction obtained in the step (P2) after the step (P2). In the step of recovering the shikimic acid, between the step (P1) and the step (P2),
The treatment solution obtained in the step (P1) is brought into contact with the strong basic anion exchange resin, and the shikimic acid contained in the treatment solution is adsorbed on the strong basic anion exchange resin, and then the shikimic acid is adsorbed. The method for producing shikimic acid according to claim 1 or 2, further comprising a step (P1a) of eluting shikimic acid from the strong basic anion exchange resin with an acidic solution. The method for producing shikimic acid according to any one of claims 1 to 3, wherein the hydrophobic synthetic adsorbent is an aromatic synthetic adsorbent. The method for producing shikimic acid according to any one of claims 1 to 4, wherein an extraction solvent containing water is used in the step (E1). The method for producing shikimic acid according to claim 3, wherein the strong basic anion exchange resin has a dimethylethanolammonium group.