JP7720049B2 - Method for decomposing amide compounds and method for producing amines - Google Patents

Description

The present invention relates to a method for decomposing amide compounds and a method for producing amines.

An example of an amine with a glycosidic bond is psychosine. Psychosine exhibits physiological activity and is thought to be involved in the pathogenesis of, for example, metabolic enzyme deficiency.

The production of amines with glycosidic bonds is important from the perspective of obtaining research reagents for clarifying the physiological activity of such amines with glycosidic bonds.

As a method for obtaining amines having glycosidic bonds, for example, Patent Document 1 discloses a method for extracting psychosine tartrate from bovine brain tissue.

Japanese Unexamined Patent Publication No. 2-200691

Amide compounds obtained by dehydration condensation between the amino group of an amine having a glycosidic bond and the carboxyl group of a fatty acid are widely distributed in nature and are contained in the flowers, leaves, roots, stems, fruits, seeds, etc. of various plants. Therefore, if such amide compounds could be decomposed and recovered as amines having glycosidic bonds, it would be possible to produce amines having glycosidic bonds from a variety of raw materials.

One possible method for decomposing an amide compound and recovering it as an amine having a glycosidic bond is to hydrolyze the amide bond in the amide compound. However, according to the inventors' investigations, when an amide compound having a glycosidic bond is hydrolyzed, the hydrolysis reaction of the glycosidic bond proceeds, and the desired amine having a glycosidic bond cannot be obtained.

The present invention was made in consideration of the above-mentioned problems, and aims to provide a method for decomposing amide compounds that can efficiently produce amines having glycosidic bonds by decomposing amide compounds having glycosidic bonds, and a method for producing amines that can produce amines having amide bonds in high yield.

To achieve the above object, the present invention provides a method for decomposing an amide compound, which includes a decomposition step in which a reaction solution containing a raw material containing an amide compound having a glycosidic bond and water is hydrothermally treated to decompose the amide compound and produce an amine having a glycosidic bond, and the pH of the reaction solution is 8.0 or higher.

The above method allows for the efficient production of amines having glycosidic bonds by decomposing amide compounds having glycosidic bonds.

The present inventors have found that when an amide compound having a glycosidic bond is decomposed by hydrothermal treatment, the hydrolysis reaction of the glycosidic bond progresses, resulting in a decrease in the yield of an amine having a glycosidic bond. For example, when glucosylceramide is used as the amide compound having a glycosidic bond, the hydrolysis reaction of the glycosidic bond progresses, resulting in the by-product of free ceramide. To solve this problem, the present inventors conducted extensive research and found that by setting the pH of the reaction solution to 8.0 or higher, the hydrolysis reaction of the glycosidic bond is suppressed and the hydrolysis reaction of the amide bond in the amide compound is promoted. As a result, an amide compound having a glycosidic bond can be decomposed to efficiently produce an amine having a glycosidic bond.

In the above method, the amide compound may contain a glycosphingolipid. According to the above method, glycosphingolipids can be decomposed particularly efficiently.

In the above method, the amide compound may include glucosylceramide. According to the above method, glucosylceramide can be decomposed particularly efficiently.

In the above method, the raw material may be a plant material.

In the above method, the content of the amide compound in the reaction solution may be 0.05% by mass or more, based on the total amount of the reaction solution. By having the mass proportion of the amide compound be 0.05% by mass or more, the amide compound can be decomposed particularly efficiently.

In the above method, the hydrothermal treatment may be carried out at a temperature of 110 to 300°C. A temperature within this range can further promote the decomposition of the amide compound.

The present invention also provides a method for producing an amine, comprising: a decomposition step in which an amide compound is decomposed by the method of the present invention; and an extraction step in which an amine having a glycosidic bond is extracted from the decomposition product obtained in the decomposition step. This production method enables the production of an amine having a glycosidic bond in high yield.

The present invention provides a method for decomposing amide compounds that can efficiently produce amines having glycosidic bonds by decomposing amide compounds having glycosidic bonds, and a method for producing amines that can produce amines having amide bonds in high yield.

The present invention will be described in detail below based on preferred embodiments. However, the present invention is not limited to the following embodiments.

In this specification, numerical ranges indicated using "to" indicate ranges that include the numerical values before and after "to" as the minimum and maximum values, respectively. In numerical ranges described in stages in this specification, the upper or lower limit of a numerical range in one stage can be arbitrarily combined with the upper or lower limit of a numerical range in another stage. In numerical ranges described in this specification, the upper or lower limit of that numerical range may be replaced with a value shown in the examples. "A or B" may include either A or B, or may include both. Unless otherwise specified, the materials exemplified in this specification can be used alone or in combination of two or more types.

(Method for decomposing amide compounds)
The method for decomposing an amide compound according to this embodiment includes a decomposition step of hydrothermally treating a reaction solution containing a raw material containing an amide compound having a glycosidic bond (hereinafter also simply referred to as "amide compound") and water to decompose the amide compound to produce an amine having a glycosidic bond. The pH of the reaction solution is 8.0 or higher.

An amide compound is a hydrophilic compound in which an amino group (-NH ₂ ) in an amine having a glycosidic bond is bonded to a carboxyl group (-COOH) of a fatty acid via an amide bond (-CONH-). Examples of amide compounds used in the present invention include, but are not limited to, glycosphingolipids.

Glycosphingolipids are composed of sugars and sphingosine bound together by glycosidic bonds. Examples of glycosphingolipids include glucosylceramide.

The sugar that serves as the base for the amide compound is not particularly limited. Examples include known sugars that can form the above-mentioned amide compounds.

The raw material to be subjected to the amide compound decomposition treatment may contain components other than the amide compound. Examples of other components include amines having glycosidic bonds, water-soluble dietary fiber, sparingly soluble dietary fiber, sugars, proteins, organic acids, etc. The content of the amide compound in the raw material is preferably 0.1% by mass or more, more preferably 0.25 to 30% by mass, and even more preferably 0.5 to 15% by mass, based on the total solid content of the raw material. If the raw material further contains an amine having a glycosidic bond, the content of the amide compound is preferably 0.25 parts by mass or more, more preferably 0.5 to 100 parts by mass, and even more preferably 5 to 50 parts by mass, per part by mass of the amine content.

Examples of raw materials include plants and seaweed. Specifically, the flowers, leaves, roots, stems, fruits, and seeds of plants and seaweed can be used. Examples of plants include legumes such as soybeans, cucurbits such as melons, grasses such as rice and corn, amaranthaceae plants such as beets, beeches such as chestnuts, Asteraceae plants such as sunflowers, and Rosaceae plants such as peaches. Because the method for decomposing amide compounds according to this embodiment decomposes amide compounds, raw materials with a low content of amines having glycosidic bonds can be used as raw materials.

The reaction liquid may contain a solvent other than water. Examples of solvents other than water include alcohol, dimethylformamide, and dimethyl sulfoxide. When the reaction liquid contains a solvent other than water, the proportion of water in the water and solvent other than water may be 5% by mass or more.

Hydrothermal treatment can be carried out by sealing the raw materials together with water in a pressure-resistant sealed container and heating it at a temperature exceeding 100°C while it is still sealed. Heating the reaction solution containing the raw materials and water in the sealed container creates a heated and pressurized environment inside the container, and hydrothermal treatment (hydrothermal synthesis) is carried out using subcritical water. The hydrothermal treatment may be carried out while stirring the reaction solution. Any known container that can be used for hydrothermal treatment can be used as the pressure-resistant sealed container without any particular restrictions. To achieve high decomposition efficiency, the filling rate of the reaction solution in the sealed container is preferably 20% by volume or more, and more preferably 40 to 80% by volume, based on the volume of the sealed container.

The content of the amide compound in the reaction solution is not particularly limited, but is preferably 0.05% by mass or more, more preferably 5% by mass or more, and preferably 25% by mass or less, and more preferably 15% by mass or less, based on the total amount of the reaction solution. When the content of the amide compound in the reaction solution is within the above range, the decomposition of the amide compound can be carried out efficiently.

The reaction conditions for the hydrothermal treatment are not particularly limited, but can be, for example, 110 to 300°C and 0.5 to 20 hours. The reaction temperature is preferably 120 to 200°C, and more preferably 140 to 195°C. A reaction temperature of 110°C or higher tends to facilitate the hydrolysis reaction of the amide bond, while a temperature of 300°C or lower tends to inhibit carbonization of the raw materials and amines having glycosidic bonds, resulting in a higher yield. The reaction time is preferably 0.2 to 20 hours, and more preferably 0.5 to 10 hours. A reaction time of 0.2 hours or longer tends to facilitate the reaction, while a reaction time of 20 hours or less tends to provide a good balance between reaction progress and costs.

The pressure inside the container during hydrothermal treatment may be equal to or higher than the saturated vapor pressure corresponding to the above-mentioned reaction temperature, but from the perspective of the pressure resistance of the equipment, saturated vapor pressure is preferable. When supplying water vapor into the sealed container, it is preferable to supply saturated water vapor at the above-mentioned reaction temperature. The pressure inside the sealed container during hydrothermal treatment can be, for example, 0.2 to 1.6 MPa.

The pH of the reaction solution is 8.0 or higher, and is preferably 9.0 or higher, more preferably 10.0 or higher, and even more preferably 11.0 or higher, in order to more efficiently decompose the amide compound and produce an amine having a glycosidic bond. The pH of the reaction solution may be 13.5 or lower.
That's fine.

The pH of the reaction solution can be adjusted by dissolving an acid or base in an aqueous solution of raw material powder containing a specific amide compound. It is preferable to use acids and bases approved as food additives. Examples of acids include citric acid, acetic acid, and malic acid. Examples of bases include sodium hydroxide.

By performing hydrothermal treatment under the above conditions, amide compounds can be decomposed to efficiently produce amines with glycosidic bonds.

(Method for producing amines having glycosidic bonds)
The method for producing an amine according to this embodiment includes a decomposition step of decomposing an amide compound and an extraction step of extracting an amine having a glycosidic bond from the decomposition product obtained in the decomposition step. The decomposition step is a step of decomposing an amide compound by the method for decomposing an amide compound according to this embodiment described above.

In the extraction process, amines having glycosidic bonds are extracted from the degradation products obtained in the degradation process. In addition to amines having glycosidic bonds, the degradation products contain sugars, amide compounds that remain undegraded, water-soluble and sparingly soluble cellulose, and their degradation products. Amines having glycosidic bonds can be extracted, for example, with ethanol heated to 60°C.

The present invention will be explained in more detail below based on examples and comparative examples, but the present invention is not limited to the following examples.

<Manufacturing Psychosine>
Example 1
3 g of Nippon Ceramide CP (trade name, manufactured by Nippon Flour Mills Co., Ltd.), which has a free ceramide content of 0.5% by mass and a glucosylceramide content of 3% by mass, was dissolved/dispersed in 147 g of ultrapure water, and 0.134 g of sodium hydroxide powder (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) was further added to obtain a reaction solution. The pH of the resulting reaction solution was measured with a pH meter and found to be 12.0. This reaction solution was placed in a 200 ml Teflon (registered trademark) container. The container was then placed in a hot air circulation autoclave (manufactured by Ashida Manufacturing Co., Ltd.) with a vessel volume of 2 m ³ , and the reaction solution was hydrothermally treated at 180 ° C for 1 hour. The hydrothermal treatment was carried out by supplying saturated steam at 180 ° C from the boiler into the autoclave vessel (pressure vessel), adjusting the steam supply rate and pressure valve so that the vessel pressure was 1 MPa, which is the saturated steam pressure of water at 180 ° C. The pressure in the tank after the hydrothermal treatment was 0.9 MPa, and the temperature in the tank was 180 ° C. The autoclave was allowed to cool naturally for 10 minutes until the pressure in the tank reached 0.7 MPa and the temperature in the tank reached 165 ° C. After natural cooling, the valve was opened, and compressed air at a pressure of 1 MPa was sent into the tank using a compressor attached to the device. Since the pressure in the tank immediately after the compressed air was sent exceeded 1 MPa, compressed air was introduced into the tank while maintaining a pressure not lower than 0.75 MPa by manually opening and closing the exhaust valve, and cooling with compressed air was started. During cooling, cooling was performed while appropriately reducing the pressure in the tank so that it did not fall below the saturated water vapor pressure at that time. Two hours after the start of cooling with compressed air, the temperature of the solution fell below 100 ° C. (the saturated water vapor pressure of the aqueous dispersion fell below normal pressure (0.1 MPa)), so the tank lid was opened, the container was removed, and the container was allowed to cool naturally to room temperature (25 ° C.). After cooling, the decomposition products of the amide compound that had precipitated or adhered to the inner surface of the container were removed using a medicine spoon.

Next, the solution and solids in the container were filtered under reduced pressure using a diaphragm pump with a hydrophilized PTFE membrane filter (Omnipore 0.2 μm JG (Merck Millipore, product name)) with a 0.2 μm mesh size. The resulting solids were dried in an oven at 120°C for 5 hours to obtain a powder of the decomposition product of the amide compound. The decomposition product of the amide compound was then prepared into a 5% dispersion in ethanol, refluxed at 60°C for 1 hour, and then filtered under reduced pressure using a diaphragm pump with a hydrophilized PTFE membrane filter (Omnipore 0.2 μm JG (Merck Millipore, product name)) with a 0.2 μm mesh size. The resulting solution was vacuum dried using a diaphragm pump at 60°C to obtain 0.04 g of psychosine concentrate powder.

(Comparative Example 1)
Except for not adding sodium hydroxide when preparing the reaction solution, 0.115 g of psychosine concentrate powder was obtained in the same manner as in Example 1. The pH of the reaction solution was 7.0.

(Comparative Example 2)
The procedure of Example 1 was repeated, except that 3 g of Nippun Ceramide CP (trade name, manufactured by Nippon Flour Mills Co., Ltd.) was dissolved/dispersed in 147 g of ultrapure water, and 0.005 g of citric acid was further added to prepare the reaction solution. 0.156 g of psychosine concentrate powder was obtained. The pH of the reaction solution was 4.0.

(Comparative Example 3)
The procedure of Example 1 was repeated, except that 3 g of Nippun Ceramide CP (trade name, manufactured by Nippon Flour Mills Co., Ltd.) was dissolved/dispersed in 147 g of ultrapure water, and 0.266 g of citric acid was further added to prepare a reaction solution. 0.115 g of psychosine concentrate powder was obtained. The pH of the reaction solution was 2.6.

<Measurement of psychosine, glucosylceramide, and free ceramide contents>
(Example 1 and Comparative Examples 1 to 3)
The contents of psychosine, glucosylceramide, and free ceramide in the resulting psychosine concentrate powder were measured by the following method. First, 20 mg of psychosine concentrate powder was diluted 50 times with a chloroform:methanol (2:1) mixed solvent to obtain a solution. 0.1 mL of the resulting solution was subjected to thin-layer chromatography (TLC) to separate psychosine, glucosylceramide, and free ceramide. A 95:12 chloroform:methanol (95:12) mixed solvent was used to separate free ceramide and glucosylceramide, and a 80:20:2 chloroform:methanol:2N aqueous ammonia solution (80:20:2) mixed solvent was used to separate psychosine. The separated psychosine, glucosylceramide, and free ceramide were quantitatively analyzed by gas chromatography. Commercially available free ceramide, glucosylceramide, and psychosine standard production samples were used as standard substances, and a constituent sphingoid base was added as an internal standard substance to perform quantitative analysis using the internal standard method. The results are shown in Table 1. The contents shown in Table 1 represent the amounts (mmol) of free ceramide, glucosylceramide, and psychosine contained in 100 g of psychosine concentrate powder. The psychosine yield is also shown in Table 1. The psychosine yield is the amount of psychosine in the obtained psychosine concentrate powder divided by the amount of glucosylceramide in the raw material.

Claims (6)

a decomposition step of hydrothermally treating a reaction solution containing a raw material containing an amide compound having a glycosidic bond and water to decompose the amide compound and produce an amine having a glycosidic bond,
the pH of the reaction solution is 8.0 or higher,
the reaction solution does not contain any solvent other than water,
the content of the amide compound in the raw material is 0.25 to 30 mass% based on the total amount of solids in the raw material;
A method for decomposing an amide compound, wherein the amide compound includes a glycosphingolipid . The degradation method according to claim 1 , wherein the amide compound comprises glucosylceramide. The decomposition method according to claim 1 or 2 , wherein the raw material is a plant raw material. The decomposition method according to any one of claims 1 to 3 , wherein the content of the amide compound in the reaction solution is 0.05 mass% or more based on the total amount of the reaction solution. The decomposition method according to any one of claims 1 to 4 , wherein the hydrothermal treatment is carried out at a temperature of 110 to 300°C. a decomposition step of decomposing an amide compound by the method according to any one of claims 1 to 5 ;
an extraction step of extracting amines having glycosidic bonds from the degradation products obtained in the decomposition step;
A method for producing an amine, comprising: