JPH07118189A - Production of d-chiro-inositol - Google Patents

Abstract

PURPOSE:To provide an industrial production method capable of using also crude kasugamycin (salt) as a raw material and further of simply hydrolyzing the kasugamycin under a mild acidic condition to produce D-chiro-inositol in a high yield and in a high purity. CONSTITUTION:A method for producing D-chiro-inositol comprises allowing the aqueous solution of crude kasugamycin hydrochloride or sulfate or the aqueous solution of crude or highly pure free kasugamycin base to pass through a column filled with the granules of a strongly acidic ion exchange resin (H<+> type) under heating, hydrolyzing the kasugamycin during the passage of the solution through the column to liberate the D-chiro-inositol from the kasugamycin, allowing the obtained acidic reaction solution containing the D-chiro-inositol to pass through a strongly acidic ion exchange resin column and a basic ion exchange resin column for removing impurities from the solution, and subsequently recovering the D-chiro-inositol as crystals from the elution solution containing the D-chiro-inositol.

Description

Detailed Description of the Invention

[0001]

FIELD OF THE INVENTION The present invention relates to D-kilo-inositol.
The present invention relates to an improved method for producing (D-chiro-inositol). D-Cyro-inositol has recently attracted attention as a therapeutic or prophylactic agent for non-insulin-dependent diabetes mellitus.

[0002]

2. Description of the Related Art A conventional method for producing D-kilo-inositol is a method obtained by extracting and purifying from a certain plant,
Method for obtaining kasugamycin by hydrolysis and method for chemically synthesizing [Journal of Organic Chemistry, Vol. 58, p. 2331-2
333, 1993] and the like are known. Among these methods, the method for producing D-kilo-inositol which enables production on an industrial scale is a method of hydrolyzing kasugamycin with a strong acid and a method of chemical synthesis.

As a method for hydrolyzing kasugamycin, high-purity kasugamycin purified to a level containing almost no impurities is used as a raw material and heated under strongly acidic conditions such as 2N trifluoroacetic acid or 5N hydrochloric acid. Method of hydrolyzing kasugamycin by means of cleaving kasugamycin to obtain D-kilo-inositol (see US Pat. No. 5,091,596)
It has been known.

[0004]

Various methods have been proposed for producing D-kilo-inositol as described above, but there are various difficulties in carrying out the production advantageously on an industrial scale. That is, the method of hydrolyzing kasugamycin to obtain D-kilo-inositol requires repeating or combining purification steps such as column chromatography and recrystallization to obtain high-purity kasugamycin as a raw material. The operation is complicated and the manufacturing cost is high. Further, the above-mentioned known method of hydrolyzing kasugamycin under strongly acidic conditions has many problems such as side reaction and low yield. In addition, the method for producing D-kilo-inositol by chemical synthesis requires stereospecific synthesis, resulting in low reaction yield and high production cost.

Therefore, an industrially advantageous method for producing D-kilo-inositol, which is an alternative to these production methods,
There is a demand for the development of a production method that requires few labor-intensive steps such as recrystallization and filtration of the produced D-kilo-inositol and is easy to operate.

The object of the present invention is to perform D-km-
An object of the present invention is to provide a method capable of producing inositol with high purity and high yield.

[0007]

[Means for Solving the Problems] The inventors of the present invention have made extensive studies in order to solve such problems. As a result, they succeeded in inventing an industrially extremely advantageous method for producing D-kilo-inositol.

Kasugamycin is a compound having a disaccharide consisting of an amino sugar having a very specific structure and D-kilo-inositol, which is very rare in nature, as a nucleus.

The present inventors have previously studied various methods for obtaining D-kilo-inositol, which is a constituent sugar, by a simple method in a high yield and a high purity by hydrolysis of kasugamycin with an acid. As a result, when the amino group of the amino sugar moiety of kasugamycin is bound and protected by a known amino-protecting group of the acyl group type, the amino sugar of kasugamycin is neutralized with the neutral sugar (D-kilo-inositol). It has been found that the glycoside bond of γ-amino acid can be efficiently cleaved to easily hydrolyze kasugamycin to produce D-kilo-inositol even under a weak acidic condition similar to that of hydrochloric acid in the concentration range of 0.05N to 0.5N.

As a result of further research, an aqueous solution of kasugamycin hydrochloride or sulfate was added to a strongly acidic ion exchange resin (H ⁺ type), preferably about 5 times the weight of the kasugamycin salt (weight) or more. When a strong acidic ion exchange resin (H ⁺ type) having a functional group as a functional group is mixed well, and thus kasugamycin is adsorbed and bound to the resin, kasugamycin hydrochloride or sulfuric acid is added to the solution phase of the mixture. HCl or H ₂ SO ₄ is released from the salt, and an acid corresponding to HCl or H ₂ SO ₄ at a concentration of about 0.05 N to 0.5 N is present as a whole. When kasugamycin bound to the resin is hydrolyzed under heating as it is without adding a new acid, the kasugamycin molecule is efficiently converted to D-kyl-inosine. It was also found that the syrup could be generated by cleavage.

When kasugamycin is bound to the strongly acidic ion exchange resin in this way, the acid conditions are much milder than those in the conventional method of the above-mentioned US patent in which kasugamycin is dissolved in 5N hydrochloric acid and hydrolyzed. 0.05N to 0.5N H
Since it can be hydrolyzed in the presence of an acid corresponding to Cl, the generation of impurities due to side reactions is extremely small, and it is found that D-kilo-inositol can be obtained in high purity from the hydrolysis reaction solution by a simple post-treatment operation. Was issued.

As described above, an aqueous solution of kasugamycin hydrochloride or sulfate is thoroughly mixed with a strongly acidic ion exchange resin,
When the kasugamycin bound to the strongly acidic ion-exchange resin is hydrolyzed in the presence of an aqueous solution containing an acid corresponding to 0.05N to 0.5N HCl generated at this time, it is produced as one decomposition product of kasugamycin. Amino sugars and other basic substances remain adsorbed on the strongly acidic ion exchange resin, and neutral D-kilo-inositol, which is the target product, does not bind to the strongly acidic ion exchange resin and is not adsorbed. Since it can be separated from the resin, D-
Chiro-inositol can be obtained in high yield and high purity even by a simple post-treatment operation.

After the hydrolysis reaction, almost pure D-kilo-inositol was converted to HC from the strongly acidic ion exchange resin.
Since it can be separated as an acidic reaction solution containing 1 or H ₂ SO ₄ , D-kilo-inositol can be efficiently separated from amino sugars, unreacted kasugamycin and other impurities adsorbed on the strongly acidic ion exchange resin. Was found.

Based on the above findings, in Japanese Patent Application No. 5-217926 (filed on August 11, 1993) previously filed, an aqueous solution of kasugamycin hydrochloride or sulfate is treated with a strongly acidic ion exchange resin (H ⁺ Type) granules to bind kasugamycin molecules to the strongly acidic functional groups of the strongly acidic ion exchange resin, thereby forming a reaction mixture containing the generated acid, which mixture is subjected to normal pressure or pressure. The present inventors have proposed a method for producing D-kilo-inositol, which has a step of performing hydrolysis reaction of kasugamycin by heating at room temperature.

As a result of further researches conducted by the present inventors, unlike the above-mentioned method, when the granular material of the strongly acidic ion-exchange resin was packed into a column and used, the resin column contained kasugamycin hydrochloride or sulfate. When the aqueous solution is continuously introduced and allowed to pass under warm conditions, the kasugamycin salt is bound to the strongly acidic ion exchange resin under the warm conditions to generate an acid in the column-passing solution, and kasugamycin It has been found that the hydrolysis reaction can proceed, which allows the D-kilo-inositol to be efficiently produced in the column passage liquid.

In the above-mentioned Japanese Patent Application No. 5-217926, an aqueous solution of kasugamycin hydrochloride or sulfate is introduced into a column packed with a granular material of a strongly acidic ion exchange resin (H ⁺ type) under heating conditions. To the
The reaction of binding the kasugamycin molecule to the strongly acidic functional group of the strongly acidic ion-exchange resin in the packed column, the reaction of generating an acid in the liquid passing through the resin column, and the hydrolysis of kasugamycin in the liquid through which D-kyl -A method for producing D-kilo-inositol, which comprises a step of causing a reaction for producing inositol in a flow-through of the column, and collecting an acidic reaction solution containing D-kilo-inositol flowing out from the column. Proposed by the inventors.

Therefore, it is used as a raw material in the method for producing D-kilo-inositol proposed in Japanese Patent Application No. 5-217926, which is a prior application, and is treated with a granular material of a strongly acidic ion exchange resin or a packed column thereof. When the aqueous solution of kasugamycin hydrochloride or sulfate is not an aqueous solution obtained by dissolving highly purified kasugamycin hydrochloride or sulfate in deionized water to the extent that it contains almost no impurities, at the final stage of its production process. In the process of obtaining high-purity crystals of D-kilo-inositol from the concentrate of the obtained neutral eluate containing D-kilo-inositol, some additional treatment such as recrystallization operation or repetition thereof may be required. It was acknowledged that fear would arise.

However, high-purity kasugamycin hydrochloride or sulfate, which is sufficiently suitable as a raw material for the above method, requires a laborious purification operation consisting of repeated column chromatography and recrystallization for preparation and a large cost. However, since the price is relatively high, it is unavoidable that the price of D-kilo-inositol in the final product is greatly affected.

The inventors of the present invention have a medium purity, for example, a purity of 60, which is a level commonly used as a raw material powder of kasugamycin for agricultural chemicals without using such high-purity and expensive kasugamycin hydrochloride or sulfate. % Is a medium purity,
Even when using crude kasugamycin hydrochloride or sulfate at a relatively low price, a strong acidic ion exchange resin (H ⁺ type) is used as in the method of Japanese Patent Application No. 5-217926, under mild acidic conditions. D by hydrolysis of kasugamycin at
Studies have been continued for the purpose of providing a method capable of producing -kilo-inositol easily, in high purity and in high yield.

As a result, medium-purity kasugamycin hydrochloride or sulfate having the above-mentioned degree was dissolved in deionized water, and the obtained aqueous solution was passed through a column of activated carbon or synthetic adsorbent, whereby When most of the impurities contained in the aqueous solution are removed by adsorption, a partially purified aqueous solution of kasugamycin hydrochloride or sulfate containing a small amount of impurities still obtained as an effluent from the activated carbon column or the synthetic adsorbent column is obtained. This aqueous solution is obtained by introducing it into a column packed with a strongly acidic ion-exchange resin (H ⁺ type) under warming conditions and allowing it to pass therethrough to hydrolyze kasugamycin under mildly acidic conditions to give D-kyl. −
It can be used without particular obstacles to produce inositol, except that the acidic hydrolysis reaction solution containing D-kilo-inositol leaving the packed column of the strongly acidic ion exchange resin (H ⁺ type) is As a post-treatment, in order to remove the coexisting basic substance, it is once again passed through a column of another strongly acidic ion exchange resin (H ⁺ type), and the acidic effluent discharged from the latter column is then discharged. , A strongly basic ion exchange resin (OH) for removal of residual acidic substances in the effluent.
^- requires that passed through a column of the mold), unless this way, D- km - additional recrystallization to achieve Shutoku high purity crystals of inositol, admitted that require such .

Further, according to a separate study conducted by the present inventors, the culture solution containing kasugamycin (in the form of free base) obtained by culturing a kasugamycin-producing bacterium was filtered, and the culture filtrate containing kasugamycin thus obtained was also: Preliminary purification in the step of collecting and preliminarily purifying kasugamycin consisting of a series of operations described below may yield an aqueous solution of kasugamycin hydrochloride or sulfate that has been partially purified in this way but still contains some impurities. Was found.

This prepurified aqueous solution was then passed through a column of activated carbon or synthetic adsorbent to remove most of the impurities in the aqueous solution and then purified by adsorption on activated carbon or synthetic adsorbent as such. When an aqueous solution of kasugamycin salt is introduced into and passed through a column packed with a strongly acidic ion exchange resin (H ⁺ type), this allows hydrolysis of kasugamycin under mild acidic conditions to produce D-kilo-inositol. They also found that there were no special obstacles. However, also in this case, the acidic reaction solution containing D-kilo-inositol produced from the hydrolysis reaction by flowing out from the column of the strongly acidic ion exchange resin (H ⁺ type) is
Similar to the above case, high-purity crystals of D-kilo-inositol must be obtained unless post-treatment consisting of re-treatment with a strongly acidic ion exchange resin column and treatment with a strong base ion exchange resin (OH ⁻ type) column. It was found that additional recrystallization operation was required to achieve the above-mentioned yield.

Furthermore, according to an experiment recently conducted by the present inventors, a highly pure or crude aqueous solution of kasugamycin free base containing no acid component was used as a strongly acidic ion exchange resin to functionalize a sulfonic acid group. Introduced into the packed column of granular material of the strong acid ion exchange resin (H ⁺ type) as a base,
When the column passing liquid is passed through the column while being maintained at a temperature of 50 to 100 ° C., the passing liquid passing through the column becomes acidic pH inside the column and flows out from the outlet of the column. It has been found that the aqueous solution has an acidic pH and also contains D-kilo-inositol as the hydrolysis product of kasugamycin.

From the above experimental results, as described above, an aqueous solution of kasugamycin hydrochloride or sulfate was introduced into a column packed with a strongly acidic ion exchange resin (H ⁺ type) and allowed to pass under heating conditions. Thus, when an attempt is made to carry out a reaction for producing D-kilo-inositol by hydrolysis of kasugamycin in the passing solution, a reaction occurs in which the kasugamycin molecule is bonded to the strongly acidic functional group of the resin. Released HCl (hydrogen chloride) or H ₂ SO ₄
The process of generating (sulfuric acid) in the liquid passing through the resin column occurs, and the process of generating some acid component by the resin, kasugamycin, and water contacting each other and interacting under heating conditions also occurs. It is more accurate to explain that an acidic state is created in the column flow-through mainly due to Then, it can be explained that in the liquid passing through the column, the reaction in which kasugamycin is hydrolyzed to produce D-kilo-inositol proceeds under the action of the mild acidic condition thus created and the strongly acidic ion exchange resin. .

Therefore, according to the first aspect of the present invention, a first step of introducing an aqueous solution of kasugamycin hydrochloride or sulfate, which is partially purified but still contains impurities, into a column of activated carbon or a synthetic adsorbent to allow the aqueous solution to pass therethrough; The kasugamycin hydrochloride or sulfate aqueous solution containing impurities obtained from the 1st step was introduced into a packed column of granular material of a strongly acidic ion exchange resin (H ⁺ type) and allowed to pass under heating conditions, whereby the kasugamycin molecule was removed. Reaction of binding in the form of an addition salt to the strongly acidic functional group of the strongly acidic ion exchange resin in a packed column of the strongly acidic ion exchange resin (H ⁺ type), and HCl or H released from kasugamycin accompanying this binding reaction. ₂ SO ₄
To produce an acid component by the mutual contact between the strongly acidic ion exchange resin, kasugamycin, and water under heating conditions. The reaction that creates an acidic state corresponding to hydrochloric acid with a concentration of 0.05 N to 0.5 N in the liquid and the passage while passing through the column while the passing liquid is in contact with the strongly acidic ion exchange resin bound with kasugamycin A column of the strongly acidic ion exchange resin and the resin is used in parallel with the reaction of hydrolyzing kasugamycin under the action of the acid generated in the liquid and the ionic resin to produce D-kilo-inositol in the passing liquid. D-kilo-inositol and HCl or H ₂ SO that are generated in the flow-through of the column and flow out from the column.
₄ , a second step of obtaining an acidic reaction solution containing ₄ and other acid components and impurities, and optionally unreacted kasugamycin, and an acidic reaction solution obtained from the second step with a strongly acidic ion exchange resin ( H ⁺ type) packed column,
This removes the basic impurities present in the reaction solution and also removes the unreacted kasugamycin that may be present in some cases, and partially purifies it as the effluent from the packed column of the strongly acidic ion exchange resin. HCI or H
₂ SO ₄ and the other acid component D- km - third step and the acidic aqueous solution obtained from the third step basic ion exchange resin Shutoku the acidic aqueous solution containing inositol (OH ^-
Type) to bind the acidic substance contained in the aqueous solution to the resin, thereby obtaining a neutralized eluate containing D-kilo-inositol exiting the column. And a fifth step of further concentrating the eluate and precipitating high-purity crystals of D-kilo-inositol from the concentrated liquid.

Next, the method of the first present invention will be specifically described.

The aqueous solution of kasugamycin hydrochloride or sulfate used as a starting material in the method of the first aspect of the present invention is a culture obtained by culturing a kasugamycin-producing bacterium improved so as to increase its production titer by a usual method. It can be an aqueous solution of kasugamycin hydrochloride or sulphate dissolved in water, which is prepared from kasugamycin separated from the culture filtrate by a usual collecting method after filtering the liquid. This kasugamycin hydrochloride or sulfate is used after it has been treated in several purification steps.For example, kasugamycin hydrochloride or sulfate produced in Japan or overseas as a kasugamycin drug substance for the production of pesticides for controlling plant diseases. It is advisable to dissolve the salt base powder in ion-exchanged water or the like before use.

The method of the first aspect of the present invention is roughly divided into 5
It consists of two processes. In the first step, kasugamycin hydrochloride or sulfate of moderate purity which can be used as a raw material was dissolved in deionized water, but an aqueous solution of kasugamycin salt containing some impurities was adsorbed and removed using a packed column of activated carbon or a synthetic adsorbent. This is a step of partial purification to some extent. The second step is the strongly acidic ion exchange resin (H
⁽⁺ Type) column is a step of passing an aqueous solution of kasugamycin salt partially purified in the first step under heating conditions to generate D-kilo-inositol by the hydrolysis reaction of kasugamycin occurring in the column. . In the third step, an acidic reaction solution containing the D-kilo-inositol produced in the second step but a small amount of basic impurities contained in the raw material and a by-product of hydrolysis is used as a strong acidic ion exchange resin. It is a partial purification step of an acidic reaction solution containing D-kilo-inositol by passing through a column.

In the fourth step, the partially purified D-kilo-inositol-containing acidic solution obtained in the third step is treated with a strongly basic ion exchange resin (OH) to remove acidic substances.
^- a step of neutralization and additional purification by that through a column of the mold). The fifth step is a step of recovering D-kilo-inositol in a highly pure crystalline form from the neutralized eluate of D-kilo-inositol obtained in the fourth step and subjected to the additional purification and neutralization. Is.

In the first aspect of the present invention, by handling all of the substances in these five steps in the state of an aqueous solution, the individual steps can be connected continuously or continuously, and the substances can be used even once. It is also possible to produce high-purity D-kilo-inositol without taking it out of the system.

Further, each step in the method of the first present invention will be described in detail.

In the first step, as a starting material, a crude aqueous solution of kasugamycin hydrochloride or sulfate, such as an aqueous solution of kasugamycin hydrochloride raw material, etc. dissolved in water such as ion-exchanged water, is used. , Packed columns of activated carbon such as activated carbon for chromatography (manufactured by Wako Pure Chemical Industries, Ltd.) or synthetic adsorbents such as SEBABEDS SP206 (manufactured by Mitsubishi Kasei Co., Ltd.) and Amberlite XAD-2 (manufactured by Rohm and Haas) Introduce and pass through.

As a result, a considerable amount of impurities in the raw material aqueous solution can be efficiently removed. Preferably, activated carbon is used and the volume of activated carbon is 1 to 200 to 400 m.
About 3 to 10 times the volume of the aqueous solution of kasugamycin salt adjusted to a concentration of kasugamycin of about 4 to 1/8 vol /
hr. It is possible to obtain an aqueous solution of kasugamycin salt from which impurities have been removed to a sufficient extent suitable for the hydrolysis reaction in the next second step by passing through the column at a rate of.

At this time, if necessary, the kasugamycin salt aqueous solution may be dissolved by heating at 70 to 80 ° C., and the kasugamycin salt aqueous solution may be passed through the adsorbent column at a high temperature without any problem. The treatment operation with this adsorbent column can significantly remove impurities without reducing the concentration of kasugamycin, thereby hydrolyzing D-kilo-inositol in the subsequent second step and subsequent steps. It is possible to prepare an aqueous solution of kasugamycin salt that has been purified to such an extent that it can be used without any trouble due to the influence of impurities in the reaction and purification post-treatment.

High-purity kasugamycin or an acid addition salt thereof containing almost no impurities, which is required as a raw material in the method of the above-mentioned US Pat. No. 5,091,596, is used for purification of crude kasugamycin. Although it is expensive because it requires various operations such as crystallization and filtration, it is expensive, but in the method of the first aspect of the present invention, the above-mentioned first step is performed, so that the medium purity is high. Since the kasugamycin salt can be used, it is possible to save a lot of labor and cost.

The "partially purified aqueous solution of kasugamycin hydrochloride or sulfate containing impurities" used as a raw material in the first step of the method of the first aspect of the present invention is obtained by culturing a kasugamycin-producing bacterium. The kasugamycin-containing culture solution was filtered, and the obtained culture filtrate was subjected to the following kasugamycin collecting step and preliminary purification step,
It is possible to obtain an aqueous solution of kasugamycin salt containing impurities in an amount corresponding to an aqueous solution obtained by dissolving the raw material of medium-purity kasugamycin hydrochloride or sulfate for agricultural chemicals in water, and use this.

That is, for example, a culture solution obtained by culturing a kasugamycin-producing bacterium is filtered, and the obtained culture filtrate is first subjected to a strongly acidic ion exchange resin (H ⁺ type) such as Diaion SK116 (manufactured by Mitsubishi Kasei). Duolite C-20
(Sumitomo Chemical Co., Ltd.), Amberlite IR124 (manufactured by Organo) or the like is passed through a column of a strongly acidic ion exchange resin (H ⁺ type) having a sulfonic acid group as a functional group to adsorb kasugamycin to the resin of the column, and The resin in the column is washed with deionized water, and then kasugamycin is desorbed and eluted from the resin through an aqueous alkali metal hydroxide solution, for example, an aqueous NaOH solution, and the eluate that has flowed out is collected by fractionation to obtain alkaline kasugamycin. The fractions were collected, and the collected alkaline kasugamycin fraction was passed through a column of a strongly acidic ion exchange resin (H ⁺ type) again, but at this time,
Although the adsorption of the kasugamycin component by the resin does not substantially occur, the excess alkali content in the alkaline kasugamycin fraction is removed by passing through at a high liquid flow rate that causes the binding and neutralization of the alkali metal hydroxide by the resin. The neutralized aqueous solution of kasugamycin from the column is concentrated and concentrated by, for example, a reverse osmosis membrane or an evaporator, and hydrochloric acid or sulfuric acid is added to the resulting concentrated aqueous solution of kasugamycin to react with kasugamycin. The aqueous solution of crude kasugamycin salt prepared by the step of recovering and prepurifying kasugamycin, which comprises obtaining an aqueous solution of kasugamycin hydrochloride or sulfate containing some residual impurities. First
It can be used as a raw material aqueous solution for the process.

In the second step of the first method of the present invention, a reaction for producing D-kilo-inositol by hydrolysis of kasugamycin is carried out in a column of a strongly acidic ion exchange resin (H ⁺ type).

The strongly acidic ion-exchange resin used in the second step is a commercially available product packed in a column, treated with an inorganic acid to prepare an H ⁺ type, and then used. The inorganic acid used is hydrochloric acid, sulfuric acid having a concentration of 1N to 5N,
There are inorganic acids such as phosphoric acid. The strong acid ion exchange resin used is preferably an ion exchange resin having a sulfonic acid group as a functional group, and commercially available strong acid ion exchange resins such as Diaion SK116 and Diaion P.
K228 (Made by Mitsubishi Kasei), Amberlite IR120
B, Amber Light 200C, Amber Light 201B
(Manufactured by Organo), Duolite C-20, Duolite C-264, Duolite XE-636 (manufactured by Sumitomo Chemical Co., Ltd.), Dowex 50W (manufactured by Muromachi Chemical Co., Ltd.) and the like.

A strongly acidic ion exchange resin column thus prepared in the H ⁺ form was heated with an aqueous solution containing kasugamycin at a concentration of 200 to 400 mg / ml by converting kasugamycin hydrochloride or sulfate into kasugamycin free base. It is introduced under the conditions and the liquid is circulated through the column.

The second step is carried out under heating conditions. Since the product D-kilo-inositol has high thermal stability, the higher the reaction temperature and the longer the reaction time, the higher the hydrolysis rate of kasugamycin. Although there are some differences in reaction efficiency depending on the type of resin used, a strongly acidic ion exchange resin (H ⁺ type) such as DIAION SK116 is used.
(Manufactured by Mitsubishi Kasei Co., Ltd.) with a strongly acidic ion exchange resin (H ⁺
Mold), the temperature in this resin column is 90
When the flow rate of the passing liquid is adjusted so that the residence time of the passing liquid in the column is 6 hours or more while being kept at ˜98 ° C., kasugamycin undergoes a hydrolysis reaction at a reaction rate of nearly 100%.

Generally, a strongly acidic ion exchange resin (H ⁺ type) which undergoes the kasugamycin hydrolysis reaction in the second step
The temperature inside the column is 50 ℃ or more under normal pressure, but 100 ℃
The following temperatures, preferably 60 to 98 ° C., particularly preferably 90 to 98 ° C., or 100 to 150 ° C. under pressurized conditions are preferably maintained. The hydrolysis of kasugamycin can be carried out at elevated temperatures. In addition, in these cases, the hydrolysis reaction in the second step requires a retention time of 6 to 6 for the passing liquid in the resin column.
It can be carried out for 40 hours, preferably 10 to 15 hours.

The hydrolysis reaction of kasugamycin is
Under pressure sufficient to suppress the boiling or the like of the water in the resin column, for example, a gauge pressure pressure of 0.1~3kg / cm ^2, preferably under a pressure of 1~ 1.2kg / cm ² 100~15
The residence time of the passing liquid in the resin column is 1.0 to
It can be carried out for 10 hours, preferably 3 to 5 hours.

When the hydrolysis reaction of kasugamycin is carried out under pressure, it is preferable that the container for accommodating the strongly acidic ion exchange resin column is a pressure resistant container and the aqueous solution to be fed therein is under pressure.

Further, the second step is repeated, that is, the acidic reaction solution flowing out from the outlet of the column of the strongly acidic ion exchange resin is recirculated to the inlet of the same column, introduced and passed through the column. It is also possible to increase the reaction yield of hydrolysis. In addition, after closing the outlet of the column, the aqueous solution of kasugamycin was temporarily stagnated in the column and reacted for a certain period of time, after confirming the production concentration of D-kilo-inositol in the solution in the column, deionized water, etc. May be fed through the column inlet to let the reaction liquid flow out from the column. In any case, during this reaction, the operation proceeded without taking the reaction solution out of the system, and the acidic reaction solution flowing out from the column was directly used as the reaction solution in which the reaction was completed.
Can be sent to the process.

In the third step of the first method of the present invention, the acidic reaction solution containing D-kilo-inositol obtained in the second step is purified by a strongly acidic ion exchange resin column for removal of basic substances. I do. In this third step, basic impurities that could not be removed in the first step and unreacted kasugamycin mixed in the acidic reaction solution obtained in the second step, or side reactions during the hydrolysis reaction in the second step, etc. It is carried out in order to remove as much as possible the basic impurities generated in 1.

The fourth step of the first method of the present invention is carried out for the purpose of neutralizing the acid generated in the reaction solution in the second step. Usually, it is carried out by passing the acidic effluent from the column after the third step through a column packed with a strongly basic ion exchange resin. The column packing amount of the resin used in the third and fourth steps varies depending on the amount of impurities contained in the hydrolysis reaction solution that has undergone the second step. It is sufficient to use approximately equal amounts of strong acid ion exchange resin and strong basic ion exchange resin, respectively. If necessary, the neutralized eluate obtained in the fourth step may be further passed through a column packed with an adsorbent such as activated carbon.

By the third and fourth steps of the first method of the present invention, impurities contained in the crude kasugamycin used as a raw material and the impurities are produced by the acid hydrolysis reaction in the second step. Most of the by-products and the by-products generated by the side reaction decomposition of kasugamycin can be removed, and D-kilo-inositol having a higher purity than crude kasugamycin can be obtained.

The basic ion-exchange resin used in the fourth step is prepared by packing a predetermined amount of resin in a column and preliminarily preparing it with an alkaline aqueous solution such as 1 to 3 N sodium hydroxide which is several times as much as this resin. It was processed to prepare an OH ^- type.
The basic ion exchange resin (OH ⁻ type) used is an ion exchange resin containing a quaternary ammonium group as a functional group, for example, Duolite A-113PLUS (manufactured by Sumitomo Chemical Co., Ltd.) and Amberlite IRA 410 (Organo Corporation). Manufactured) and the like.

These first to fourth steps can be carried out independently and at different times, but some or all of the columns used in these steps are connected by conduits and connected. By adjusting the flow rate of the solution through each column by the control of a pump inserted in the conduit,
Part or all of the first step to the fourth step can be performed continuously.

In this case, the flow rate of the solution to be treated in each column is defined by the column flow rate in the second step in which the hydrolysis reaction is carried out, and in order to increase the hydrolysis rate in the second step. For this, it is also an effective means to make pressurized conditions in the column in the second step and heat the solution during the reaction to 100 ° C. or higher. In addition, since the resins used in the second to fourth steps can be regenerated by acid or alkali treatment, if they are regenerated while once packed in the column, they can be reused and repeatedly used semipermanently.

In the fifth step of the first method of the present invention, the D-kilo-inositol obtained from the strongly basic ion exchange resin column used in the fourth step is contained but substantially free of impurities. D-Kilo-inositol is recovered from the combined eluate. For this recovery, the eluate described above is concentrated, and high-purity D-kilo-inositol crystals are precipitated from the concentrate, and ethanol may be added to the concentrate for the precipitation of the crystals. In this case, generally, the usual and conventional means for precipitating sugar crystals from a concentrated aqueous solution containing sugar can be applied.

As described earlier in this specification, even in high purity,
When a crude aqueous solution of kasugamycin free base is passed through a column packed with a strongly acidic ion exchange resin (H + type) having a sulfonic acid group as a functional group at a temperature of 50 to 100 ° C., the liquid passing through the column is acidic. The pH of the solution becomes pH, and kasugamycin is hydrolyzed under the action of acidic conditions and strongly acidic ion-exchange resin in this passing solution to give D-kil-
It was found from the experiment that a reaction for producing inositol occurred and an acidic reaction solution containing D-kilo-inositol could be obtained as an effluent from the outlet of the column.

Accordingly, the first aspect of the present invention can be modified to produce D-kilo-inositol using as raw material an aqueous solution of crude or high-purity kasugamycin free base.

Therefore, according to the second aspect of the present invention, the first step and the first step in which an aqueous solution of kasugamycin free base partially purified but still containing impurities is introduced into a column of activated carbon or a synthetic adsorbent and allowed to pass therethrough. An aqueous solution of kasugamycin free base containing impurities obtained from
An aqueous solution of kasugamycin free base, which has been separately purified in advance to a high purity without any steps, is treated with a strongly acidic ion exchange resin (H
⁽⁺ Type) granules are introduced into a packed column and passed under heating conditions, whereby the kasugamycin molecules are allowed to pass strongly acidic functionalities of the strongly acidic ion exchange resin (H ⁺ type) packed column. The acid component is generated by the reaction of binding to a group and the mutual contact of the strongly acidic ion exchange resin, kasugamycin and water under heating conditions to hydrolyze the kasugamycin under the action of the ion exchange resin to produce D. -Kilo-inositol is produced in the flow-through of the column of the strongly acidic ion-exchange resin, and the D-kilo-inositol and the acid component flowing out from the column are unreacted with impurities which may be mixed. also a second step of Shutoku reaction solution acidic containing kasugamycin, through the reaction solution acidic obtained from the second step to a packed column of a strongly acidic ion exchange resin (H ⁺ type), which It further removes the basic impurities present in the reaction solution and also removes any unreacted kasugamycin present and is partially purified as an effluent from the packed column of the strongly acidic ion exchange resin. And a third step of obtaining an acidic aqueous solution containing an acid component and D-kilo-inositol, and the acidic aqueous solution obtained from the third step is passed through a column packed with a basic ion exchange resin (OH ⁻ type). The fourth step of binding the acidic substance contained in the aqueous solution to the resin by the method to obtain a neutralized eluate containing D-kilo-inositol exiting from the column, and further concentrating the eluate and The fifth step of precipitating high-purity D-kilo-inositol crystals from the concentrated liquid is carried out, except that in the second step, kasugamycin release which is separately purified to high purity without passing through the first step is performed. When using an aqueous solution of the base is characterized by omitting the first step, D- km - method for producing inositol is provided.

The aqueous solution of partially purified but still impure kasugamycin free base used to introduce and pass through the column of activated carbon or synthetic adsorbent in the first step of the second process of the present invention is kasugamycin. The culture solution obtained by culturing the producing bacterium is filtered, and the obtained culture filtrate is first passed through a column of a strongly acidic ion exchange resin (H ⁺ type) to adsorb kasugamycin to the resin of the column, and then the column Resin is washed with deionized water, and then kasugamycin is desorbed from the resin through an aqueous alkali metal hydroxide solution to elute, and the alkaline kasugamycin fraction is collected by fractionation of the alkaline eluent thus obtained,
The collected kasugamycin fraction was treated with a strongly acidic ion exchange resin (H
⁺ Type) column, but at such a high flow rate as to cause binding and neutralization of the alkali metal hydroxide by the resin, although adsorption of the kasugamycin component by the resin does not occur substantially. Recovery and pre-purification of kasugamycin, which comprises neutralizing the kasugamycin fraction through and concentrating the neutralized kasugamycin aqueous solution exiting the column, thereby collecting an aqueous solution of kasugamycin free base with some residual impurities Can be obtained at the stage of.

Furthermore, the temperature in the column of the strongly acidic ion exchange resin used for carrying out the hydrolysis reaction of kasugamycin in the second step of the second method of the present invention is 50 ° C. under normal pressure.
However, the temperature is 100 ° C. or lower, preferably 60 to 98.
Maintained at a temperature of ℃ or under pressure condition 100 ~
It can be maintained at a temperature of 150 ° C.

The second method of the present invention is different in that the kasugamycin free base is used in place of the kasugamycin hydrochloride or the sulfate used in the first invention, but the second method is used.
The activated carbon or synthetic adsorbent used in the first step of the method of the present invention and the various ion exchange resins used in the second to fourth steps are respectively the adsorbent used in the first step of the first method of the present invention. It can be the same as various ion exchange resins used in the second to fourth steps.

The first to fifth steps of the second method of the present invention can be carried out by operating in the same manner as the first to fifth steps of the first method of the present invention.

Furthermore, the present inventors have provided a second method of the second invention.
A series of experiments were carried out to carry out the process, but a high-purity or crude aqueous solution of kasugamycin free base was passed through a packed column of a strongly acidic ion exchange resin (H ⁺ type) having a sulfonic acid group as a functional group. When the temperature is in the range of 50 to 150 ° C under normal pressure or under pressure, the concentration of kasugamycin in the aqueous solution charged in the column, the flow rate of the passing liquid, the amount of resin in the column, other operating factors, etc. It was also found that the adjustment reaction to the respective factors allows the production reaction of D-kilo-inositol to proceed by hydrolysis of kasugamycin.

Therefore, according to the third aspect of the present invention, an aqueous solution of kasugamycin free base partially purified but still containing a very small amount of impurities or highly purified pre-purified kasugamycin free base is treated with a sulfonic acid group. The strong acid ion exchange resin (H ⁺ type) as a base is introduced into a packed column and passed through the column at a temperature in the range of 50 to 150 ° C., whereby the kasugamycin molecules are allowed to pass through the strongly acidic ion exchange resin (H ⁺ type). ⁺ Type) to generate an acid component by the reaction of binding to the sulfonic acid functional group of the strongly acidic ion exchange resin in a packed column and the mutual contact of the strongly acidic ion exchange resin, kasugamycin, and water under heating conditions. Then, kasugamycin is hydrolyzed under the action of the ion exchange resin to produce D-kilo-inositol in the column of the strongly acidic ion exchange resin, and further, D-Kilo-flowing out of the column
There is provided a method for producing D-kilo-inositol, which comprises collecting an acidic reaction solution containing inositol and unreacted kasugamycin.

This third method of the present invention comprises the first and second methods.
Can be operated in the same manner as in the second step of the method of the present invention.

An apparatus suitable for continuously carrying out the first to fourth steps of the first and second inventive methods is shown schematically in FIG. 1 of the accompanying drawings.

This apparatus comprises a packed column 1 of activated carbon and a strongly acidic ion exchange resin (H ⁺ type) which functions as a hydrolysis reaction tower.
Packed column 2, a strongly acidic ion exchange resin (H ⁺
Type) packed column 3 and a strong basic ion exchange resin (OH ⁻ type) packed column 4 for purification.

At the inlet of the top of the activated carbon column 1, a supply pipe 7 for an aqueous solution of crude kasugamycin (salt) used as a raw material is provided. The outlet at the bottom of the column 1 is connected to the inlet of the strongly acidic ion-exchange resin column 2 for hydrolysis reaction by a conduit 8, and this conduit 8 has an opening / closing valve on the outlet side of the column 1 and the inlet side of the column 2. 12 and the on-off valve 13 are inserted, and the liquid feed pump 5 is inserted at an intermediate position between the valves 12 and 13. Pump 5 is preferably a type of liquid flow rate controllable pump capable of controlling the flow rate of liquid through conduit 8.

The outlet of the strongly acidic ion exchange resin column 2 (hydrolysis reaction column) and the inlet of the strongly acidic ion exchange resin column 3 for purification are connected by a conduit 9, and the conduit 9 flows out from the column 2. A degasser (valve) 6 for removing the gas to the outside when the gas is mixed in the reaction solution is also installed, and an on-off valve 14 is also installed.

The outlet of the strongly acidic ion exchange resin column 3 for purification and the inlet of the strongly acidic ion exchange resin column 4 for purification are connected by a conduit 10 having an on-off valve 15. The outlet of the column 4 is provided with a delivery pipe 11 for the solution coming out of the column 4. The delivery pipe 11 is further arranged to deliver the neutralized and purified D-kilo-inositol aqueous solution leaving the column 4 to a receiver (not shown) for collecting the aqueous solution. A concentrator (not shown) for concentrating the D-kilo-inositol aqueous solution collected in the receiver is provided behind the column 4.

Since the strongly acidic ion exchange resin column 2 as the hydrolysis reaction column heats the resin in the column and the solution in the reaction flowing through the column to a high reaction temperature of 50 ° C. or higher, A suitable heating device such as a heating jacket or oven surrounding the column is installed.

Next, the production method of the first present invention will be specifically described with reference to the following examples, but the present invention is not limited to the following examples.

Example 1 This example illustrates the preparation of D-kilo-inositol using a crude aqueous solution of kasugamycin hydrochloride as a raw material. As schematically shown in FIG. 1 of the accompanying drawings, a packed column of activated carbon, a packed column of a strongly acidic ion exchange resin (H ⁺ type) as a hydrolysis reactor, and a strongly acidic ion exchange resin (H ⁺ Type) packed column and a strong basic ion exchange resin (OH ⁻ type) packed column for purification are connected by a conduit, and the first to fourth steps of the first method of the present invention are continuously performed. An apparatus was prepared so that it could be carried out.

(Step 1) 24 g of crude raw powder of kasugamycin hydrochloride (manufactured by Nippon Kayaku Co., Ltd., 62.3% in terms of kasugamycin free base) was dissolved in 120 ml of deionized water with heating. The total amount of the obtained aqueous solution of kasugamycin hydrochloride containing impurities was placed on the top of a packed column (internal diameter: 1.5 cm, length: 14 cm) of an activated carbon column having a volume of 25 ml, and a liquid was passed through the activated carbon column by driving a pump.

In this case, 120 ml of deionized water was added to the top of the activated carbon column immediately before the aqueous solution of kasugamycin hydrochloride had cleared from the top surface. The aqueous solution was passed through the activated carbon column at a flow rate of about 6 ml / hour.

(Second step) Casugamycin hydrochloride aqueous solution (p
H 4) is a strongly acidic ion exchange resin DIAION SK1
Column (inner diameter 2.5 cm, length 24.5 cm) packed with 120 ml capacity of 16 (H ⁺ type) (manufactured by Mitsubishi Kasei Co., Ltd.)
(The column is installed in an oven at 90 ° C.), and the column residence time of the passing liquid was set to about 7 hours.

Kasugamycin was hydrolyzed in the solution passing through the strongly acidic ion exchange resin column to produce D-kilo-inositol. From the outlet of the column, D-
Acidic reaction solution containing pH-inositol (pH 1.0)
Spilled.

(Third Step) The acidic reaction solution obtained in the second step was treated with a strongly acidic ion exchange resin, Duolite C-2.
0 (H ⁺ type) (Sumitomo Chemical Co., Ltd.) 100 ml
Column packed with volume (internal diameter 2.4 cm, length 22 cm)
Was introduced into and passed through.

(Fourth step) In the third step, Duolite C-
20 acidic column aqueous solution, strong basic ion exchange resin, Duolite A-113PLUS (OH ^- type)
It was introduced into a column (internal diameter 2.4 cm, length 26.5 cm) packed with 120 ml capacity (Sumitomo Chemical Co., Ltd.) and allowed to pass.

From the outlet of the Duolite A-113 PLUS column, a neutralized eluate containing D-kilo-inositol was obtained.

The time required for the first to fourth steps carried out as described above was 40 hours in total including the washing of the column with water.

(Fifth Step) In the fourth step, Duolite A-
After the D-kilo-inositol aqueous solution was obtained from the 113PLUS column, the column was washed with water. The content of D-kilo-inositol in the mixed solution obtained by combining the washing solution with an aqueous solution of D-kilo-inositol was measured by high performance liquid chromatography to be 6.8 g (theoretical value 7.1 g) (weight yield 96%, purity It was about 99%). Furthermore, after concentrating the above mixture, ethanol was added to crystallize D-kilo-inositol to obtain 6.5 g (weight yield 92%, purity about 100%).

The thus obtained D-kilo-inositol has a specific optical rotation of [α] ^D ₂₂ + 65 ° (c 1.0,
Water), melting point 238 ° C., and these physicochemical properties showed that this substance was pure.

Next, the method of the second aspect of the present invention will be specifically described with reference to Examples 2 to 3 below, but the method is not limited to these Examples.

Example 2 This example illustrates the case of producing D-kilo-inositol using an aqueous solution of crude kasugamycin (in the form of free base) containing impurities as a raw material. Kasugamycin (free base) is a crude substance obtained by treating a Kasugamycin-containing culture solution obtained by culturing a Kasugamycin-producing bacterium in the following steps of collecting and prepurifying Kasugamycin.

(A) Production of Crude Kasugamycin from Culture Solution of Kasugamycin Producing Streptomyces kasugaensis, a Kasugamycin-producing bacterium, by a conventional method to obtain a culture solution containing kasugamycin, which was filtered to obtain kasugamycin. 2 liters of culture filtrate containing (Kasugamycin titer 9.8 mg / ml) was collected. This culture furnace liquid was passed through a column (internal diameter: 5 cm, length: 76.5 cm) packed with 1.5 l of a strongly acidic ion exchange resin, Duolite C-20 (H ⁺ type) to adsorb kasugamycin to the resin.

Next, this resin column was washed with 4.5 l of deionized water and then eluted with 4.5 l of a 2N-NaOH aqueous solution. The alkaline eluate obtained from the column was fractionated to collect an alkaline active fraction containing kasugamycin, and the active fraction was treated with 1.5 l of a strongly acidic ion exchange resin, DIAION SK116 (H ⁺ type). It was introduced into a packed column (internal diameter 5 cm, length 76.5 cm) and passed through at a flow rate which was maximized as much as possible. At this time, most of kasugamycin in the active fraction passed through without being adsorbed on Diaion SK116 resin, but since the alkaline substance was bound to the resin, the kasugamycin active fraction was neutralized. Thus, a neutralized aqueous solution of kasugamycin was obtained as an outflow from the Diaion SK116 column.

This neutralized aqueous solution of kasugamycin 23
00ml was collected, concentrated on a rotary evaporator,
75 ml of a concentrated aqueous solution containing kasugamycin (free base) at a concentration of 200 mg / ml was obtained.

By the way, when this concentrated aqueous solution was evaporated to dryness under reduced pressure, it was found that the solid residue contained kasugamycin (free base) in a content of about 60%.

(B) Production and purification of D-kilo-inositol from crude kasugamycin (free base) 75 ml of a concentrated aqueous solution (pH about 7) of kasugamycin (free base) obtained in step (A) above was carried out. It was introduced into the packed column of activated carbon used in the first step of Example 1 in the same manner as in Example 1 and passed through at a flow rate of about 6 ml / hour.

A strongly acidic ion exchange resin DIAION SK116 (as a hydrolysis reaction column used in the second step of Example 1 was prepared by using an aqueous solution of kasugamycin (free base) partially purified in an activated carbon column but still containing impurities. (H ⁺ type) packed column in the same manner as in the second step of Example 1 with a liquid temperature of 90
It was allowed to pass at about 7 hours for about 7 hours. This diamond SK
Acidic reaction solution (pH 1.4) flowing out from the 116 column
Is D-kilo produced by hydrolysis of kasugamycin.
Contains inositol.

The above acidic reaction solution was added to the third solution of Example 1.
In the same manner as in the step, it was introduced into a Duolite C-20 (H ⁺ type) column for treatment, and the effluent from the Duolite C-20 column was treated in the same manner as in the fourth step of Example 1 in the Duolite A
-113 PLUS (OH ^- type) column was introduced and processed.

When the neutralized aqueous D-kilo-inositol solution obtained as the effluent from the Duolite A-113 PLUS column was treated in the same manner as in the fifth step of Example 1, 5.2 g of D-kilo-inositol ( Purity about 99%) was obtained.

Example 3 In this example, when kasugamycin (free base) that had been highly purified in advance was used, even if the treatment with the activated carbon column in the first step of Example 1 was omitted, high-purity kasugamycin (free base) was obtained. Free base) is directly subjected to the hydrolysis reaction according to the second step of Example 1 and the resulting acidic reaction solution is then subjected to a post-treatment in the third step to the fourth step of Example 1,
It illustrates that high-purity D-kilo-inositol can be produced in high yield.

(A) Preparation of high-purity kasugamycin (free base) 18 g of crystals of kasugamycin hydrochloride were dissolved in 700 ml of ion-exchanged water, and 600 ml of strongly acidic ion-exchange resin Duolite C-20 (H ⁺ type) was dissolved. Column (inner diameter 5c
m, length 30 cm), and kasugamycin was adsorbed on the resin. Next, this column was washed with 1.8 l of ion-exchanged water and then eluted with 2.4 l of 2% aqueous ammonia to obtain an eluate containing kasugamycin in the form of a free base. At this time, when the eluate was fractionated by 100 ml, kasugamycin was separated into fraction no. It started to elute little by little from 10. These kasugamycin fractions were collected and freeze-dried to obtain 15 g (98% purity) of free base kasugamycin.

(B) Production and purification of D-kilo-inositol from high-purity kasugamycin (free base) The high-purity kasugamycin (free base) obtained in the above step (A) was dissolved in deionized water. Thus, about 70 ml of an aqueous solution of kasugamycin containing kasugamycin at a concentration of about 200 mg / ml and containing almost no impurities was prepared.

This aqueous solution was used as the strongly acidic ion exchange resin used in the second step of Example 1, Diaion SK116 (H ⁺
Type) packed column directly at a liquid temperature of 90 ° C.
In the same manner as in the second step of Example 1, kasugamycin was subjected to a hydrolysis reaction by passing it at a reaction temperature of 90 ° C. at a flow rate of about 6 ml / hour with a column residence time of about 7 hours.

The acidic reaction solution (pH 1.4) flowing out from the Diaion SK116 column was used to generate the D-
Contains kilo-inositol.

When this acidic reaction solution is treated in the same manner as in the operations of the third, fourth and fifth steps of Example 1 below,
High-purity D-kilo-inositol (purity 100%)
6.7 g (yield 96%) was obtained.

[0097]

INDUSTRIAL APPLICABILITY According to the present invention, D-kilo-inositol can be produced simply and continuously from an acid addition salt of kasugamycin or a free base under very mild conditions in high yield and high purity. Is possible. In the case of large-scale production, strong acids are not used, so a small amount of ion-exchange resin is required for neutralization, and the power consumption used for treating sewage and distilling water is greatly reduced accordingly. There is a great advantage in that the production equipment per unit yield can be small. Moreover, even impure kasugamycin is economical because it can be used as a raw material.

[Brief description of drawings]

1 is a schematic illustration of an apparatus suitable for carrying out the method of the present invention.

[Explanation of symbols]

1 activated carbon packed column 2 strongly acidic ion exchange resin (H ⁺ form) hydrolysis reactor 3 strongly acidic ion exchange resin consisting of packed column (H ⁺ form) packed column 4 strongly basic ion exchange resin (OH ^- type ) Packed column 5 Liquid feed pump 6 Degasser 8 Conduit 12 Open / close valve

 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Kiyoshi Hirasawa 3-8 Morinosato, Atsugi, Kanagawa Prefecture (72) Inventor Tomio Takeuchi 5-11 East Higashigotanda, Shinagawa-ku, Tokyo New Mansion 701

Claims (8)

[Claims] 1. A first step of introducing a partially purified aqueous solution of kasugamycin hydrochloride or a sulfate containing an impurity into a column of activated carbon or a synthetic adsorbent and passing it through, and impurities obtained from the first step. An aqueous solution containing kasugamycin hydrochloride or sulfate containing strongly acidic ion exchange resin (H
⁽⁺ Type) granules are introduced into a packed column and passed under heating conditions, whereby the kasugamycin molecules are allowed to pass strongly acidic functionalities of the strongly acidic ion exchange resin (H ⁺ type) packed column. Reaction to form a salt in the form of an addition salt, and HCl or H ₂ SO ₄ liberated from kasugamycin accompanying this binding reaction is produced in a strongly acidic ion-exchange resin column permeate and the reaction under heating conditions is performed. A reaction for producing an acid state corresponding to hydrochloric acid having a concentration of 0.05 N to 0.5 N in the passing liquid mainly due to generation of an acid component by mutual contact between strongly acidic ion exchange resin, kasugamycin and water. While passing through the column while the passing solution is in contact with a strongly acidic ion exchange resin bound with kasugamycin, kasugamycin is produced under the action of the acid generated in the passing solution and the ion exchange resin. And D-kilo-inositol are hydrolyzed to produce D-kilo-inositol in the flow-through solution in parallel with each other in the flow-through solution of the strongly acidic ion-exchange resin and the column of the resin, and the reaction solution is discharged from the column. A second step of obtaining an acidic reaction solution containing incoming D-kilo-inositol, HCl or H ₂ SO ₄ , other acid components and impurities and optionally also unreacted kasugamycin, The acidic reaction solution thus prepared is treated with a strongly acidic ion exchange resin (H
⁺ Type) packed column, thereby removing the basic impurities present in the reaction solution and optionally unreacted kasugamycin, and filling the strongly acidic ion exchange resin packed column. The third step of obtaining an acidic aqueous solution containing HCl or H ₂ SO ₄ and other acid components and D-kilo-inositol as an effluent from the third step, and the acid obtained from the third step. Of the basic ion-exchange resin (OH ^- type) is passed through the column to bind the acidic substance contained in the aqueous solution to the resin, and the neutralization containing D-kilo-inositol exiting the column is performed. The fourth step of collecting the obtained eluate, and further concentrating the eluate and producing a highly pure D-kilo-
A fifth step of precipitating crystals of inositol, the method for producing D-kilo-inositol. 2. Used for introducing into and passing through a column of activated carbon or synthetic adsorbent in the first step,
An aqueous solution of kasugamycin hydrochloride or sulfate, which is partially purified but still contains impurities, is obtained by filtering the culture solution obtained by culturing a kasugamycin-producing bacterium, and then adding the obtained culture filtrate to a strong acidic ion exchange resin (H ⁺ Type) column to adsorb kasugamycin to the resin of the column, then wash the resin of the column with deionized water, and then desorb and elute kasugamycin from the resin through an aqueous alkali metal hydroxide solution, The alkaline kasugamycin fractions were collected by fractionation of the alkaline eluate thus obtained, and the collected kasugamycin fractions were treated with a strongly acidic ion exchange resin (H ⁺ type).
However, at this time, although the adsorption of the kasugamycin component by the resin does not substantially occur, the kasugamycin is passed through at a high flow rate that causes binding and neutralization of the alkali metal hydroxide by the resin. Neutralize the fractions,
The neutralized aqueous solution of kasugamycin discharged from the column is concentrated, and hydrochloric acid or sulfuric acid is added to the obtained concentrated aqueous solution of kasugamycin to react, whereby an aqueous solution of kasugamycin hydrochloride or sulfate containing some residual impurities is collected. The method according to claim 1, which is obtained in the stage of recovery and preliminary purification of kasugamycin, which comprises 3. The temperature in the column of the strongly acidic ion exchange resin used for carrying out the hydrolysis reaction of kasugamycin in the second step is 50 ° C. or higher under normal pressure, but 100 ° C. or lower, preferably 60 to 98 ° C. To maintain the temperature of
Alternatively, the method according to claim 1, which is maintained at a temperature of 100 to 150 ° C. under pressure. 4. A column of activated carbon or synthetic adsorbent used in the first step, a packed column of a strongly acidic ion exchange resin (H ⁺ type) as a hydrolysis reaction column used in the second step,
A column of a strongly acidic ion exchange resin (H ⁺ type) as a tower for partially purifying the reaction solution used in the third step, and a strong basic ion exchange resin (O ⁺ as a neutralization tower of the reaction solution used in the fourth step).
H ^- type) packed columns are mutually connected in series by conduits, and if desired, the outlet of the column of activated carbon or synthetic adsorbent and the inlet of the column for hydrolysis reaction are connected. The method according to claim 1, wherein a liquid feed pump is inserted in the conduit. 5. A first step of introducing a partially purified aqueous solution of kasugamycin free base still containing impurities into a column of activated carbon or a synthetic adsorbent, and passing it through, and releasing kasugamycin containing impurities obtained from the first step. An aqueous solution of a base or an aqueous solution of kasugamycin free base, which was separately purified to a high purity without passing through the first step, was introduced into a column packed with a granular material of a strongly acidic ion exchange resin (H ⁺ type). Reaction under a heating condition by which the kasugamycin molecule is bonded to the strongly acidic functional group of the strongly acidic ion exchange resin in the packed column of the strongly acidic ion exchange resin (H ⁺ type). An acid component is generated by mutual contact between the strongly acidic ion exchange resin, kasugamycin and water, and kasugamycin is hydrolyzed under the action of the ion exchange resin to give D-key. - inositol was generated in the flow-through of the column of the resin, flowing out from the column D
-A second step of obtaining an acidic reaction solution containing chiro-inositol and an acid component and optionally mixed impurities and unreacted kasugamycin; and the acidic reaction solution obtained from the second step is treated with a strongly acidic ion. The mixture is passed through a packed column of exchange resin (H ⁺ type) to remove the basic impurities present in the reaction solution and also to remove unreacted kasugamycin, which may be present in the reaction solution. The third step of obtaining an acidic aqueous solution containing a partially purified acid component and D-kilo-inositol as an effluent discharged from the packed column of the resin, and the acidic aqueous solution obtained from the third step is made basic. Neutralization with D-kilo-inositol exiting the column by passing it through a packed column of ion exchange resin (OH ^- type) to bind the acidic material contained in the aqueous solution to the resin. The fourth step of collecting the obtained eluate and the fifth step of further concentrating the eluate and precipitating high-purity D-kilo-inositol crystals from the concentrated solution are carried out, provided that in the second step, A method for producing D-kilo-inositol, which comprises omitting the first step when an aqueous solution of kasugamycin free base purified separately without the first step is used. 6. Used in the first step to introduce and pass through a column of activated carbon or synthetic adsorbent,
An aqueous solution of kasugamycin free base that was partially purified but still contained impurities was obtained by filtering the culture solution obtained by culturing the kasugamycin-producing bacterium, and then adding the obtained culture filtrate to a strongly acidic ion exchange resin (H ⁺ type). Kasugamycin is adsorbed on the resin of the column through a column, and then the resin of the column is washed with deionized water, after which Kasugamycin is desorbed and eluted from the resin by passing through an aqueous alkali metal hydroxide solution. The alkaline kasugamycin fraction was collected by fractionation of the obtained alkaline eluate, and the collected kasugamycin fraction was passed through a column of a strongly acidic ion exchange resin (H ⁺ type), but at this time, adsorption of the kasugamycin component by the resin was performed. Although it does not substantially occur, the resin is passed through at such a high flow rate as to cause the binding and neutralization of the alkali metal hydroxide by the resin. A step of recovery and pre-purification of kasugamycin comprising neutralizing the isine fraction and concentrating the neutralized aqueous kasugamycin solution exiting the column, thereby collecting an aqueous solution of kasugamycin free base with some residual impurities. The method according to claim 5, which is obtained by 7. The temperature in the column of the strongly acidic ion exchange resin used for carrying out the hydrolysis reaction of kasugamycin in the second step is 50 ° C. or higher under normal pressure, but 100 ° C. or lower, preferably 60 to 98 ° C. To maintain the temperature of
Alternatively, the method according to claim 5, which is maintained at a temperature of 100 to 150 ° C under pressure. 8. A strongly acidic ion-exchange resin having a sulfonic acid group as a functional group, which is an aqueous solution of kasugamycin free base partially purified but still containing a very small amount of impurities, or kasugamycin free base highly purified in advance. (H ⁺ type)
Introduced into the packed column of the granular material of
Passing in the range of 150 ° C., which allows the kasugamycin molecules to bind to the sulfonic acid functional groups of the strongly acidic ion exchange resin (H ⁺ form) in a packed column of strongly acidic ion exchange resin (H ⁺ form) and under heating conditions. An acid component is generated by mutual contact between the strongly acidic ion exchange resin, kasugamycin and water, and kasugamycin is hydrolyzed under the action of the ion exchange resin to convert D-kilo-inositol to the strongly acidic ion exchange resin. A method for producing D-kilo-inositol, which comprises producing an acidic reaction solution containing D-kilo-inositol and unreacted kasugamycin produced in the column and flowing out from the column.