JPH0416195A - Production of methylglycoside monoester of fatty acid - Google Patents

Abstract

PURPOSE:To enable selective and efficient, obtaining of the subject monoester by reacting a methylglycoside, etc., fatty acids, etc., with a neutral thermoduric immobilized hydrolase in a specific solvent. CONSTITUTION:(A) Methylglycosides are obtained by selection from sugars prepared by containing methyl groups as an aglycone in N hemiacetal (anomeric) hydroxyl groups of 5-7C monosaccharides and disaccharides compose of hexoses. (B) Fatty acids are then obtained by selection from 6-22C (un)saturated fatty acids and esters of the aforementioned fatty acids with 1-3C lower alcohols. (C) Aneutral thermoduric immobilized hydrolase is subsequently obtained by immobilizing a yeast lipase, etc., on active carbon, etc. To 1mol ingredient (B), is added 0.05-0.5mol ingredient (A). To 100 pts.wt. ingredient (B), is added the ingredient (C) so as to provide 0.1-10000 pts.wt. amount of the ingredient (C). Reaction is then carried out in the presence of an organic solvent, such as tertiary butyl alcohol, having electrical characteristics of >=0.5 D dipole moment without containing water at 50-120 deg.C for about 24hr to produce the objective methylglycoside monoester of the fatty acids.

Description

[Detailed Description of the Invention] Top side of chestnuts, 1L number The present invention is directed to the production of methyl glycoside fatty acid esters from methyl glycoside and fatty acids by enzymatic reaction.
The present invention relates to a method for producing methyl glycoside fatty acid monoesters that can selectively synthesize monoesters.

Conventionally, as a method for producing alkyl glycoside fatty acid esters, a chemical synthesis method has been proposed in which a hexamonosaccharide glycoside is reacted with a fatty acid or a fatty acid derivative in the presence of a dehydration condensation agent or a basic compound. (U.S. Patent No. 4,716,152
Publication No.).

However, with this chemical synthesis method, it is difficult to obtain only monoesters with high selectivity, and moreover, it requires the use of large amounts of expensive dehydration condensation agents, and fatty acid derivatives (such as acid chlorides) that are difficult to use industrially. There are disadvantages such as using .

In contrast, enzymatic production methods have recently attracted attention, and a method has been proposed in which a fatty acid is allowed to act on an alkyl glycoside having an alkyl group of 03 or more in the presence of a certain amount of water or less and a fatty acid degrading enzyme. (Unexamined Japanese Patent Publication No. 2-94
Publication No. 36). Furthermore, a method has been proposed in which a hydrolase is allowed to act on a mixture of an alkyl glycoside having an alkyl group of 02 to C1 and a fatty acid or a lower alcohol ester of a fatty acid (W089101480).

However, in the former method, the use of lower alkyl glycosides in which the alkyl group is methyl or ethyl is ineffective, and in the latter method, it is ineffective in treating methyl glycosides. It is difficult to obtain alkyl glycoside fatty acid esters with high content. In particular, when the amount of fatty acid is increased relative to the alkyl glycoside in order to increase reaction efficiency, there is a drawback that a large amount of polysubstituted products such as diesters are produced as by-products.

The present invention was made in view of the above circumstances, and it is possible to suppress the by-product of polysubstituted products such as diesters, selectively obtain only monoesters, and efficiently perform ester synthesis or transesterification. The purpose of the present invention is to provide a method for producing methyl glycoside fatty acid monoesters.

In order to achieve the above object, the present inventors conducted intensive studies and found that the methyl glycoside of a sugar selected from monosaccharides having 5 to 7 carbon atoms and dimonosaccharides consisting of hexoses has a carbon number of When reacting fatty acids selected from 6 to 22 saturated and unsaturated fatty acids and esters of these fatty acids and lower alcohols having 1 to 3 carbon atoms, a neutral heat-resistant immobilized hydrolytic atom is used, and a dipole When reacted in a substantially water-free organic solvent having electrical properties with a moment of 0.5 Debye or more, the above fatty acids The ester synthesis or transesterification reaction is easily carried out efficiently and well, achieving a high reaction rate, and the selectivity of the reaction is also high, reducing the by-product of polysubstituted products such as diesters (diacyls). It was found that only monoesters (monoacyls) were preferentially given. In addition, in this case, even if fatty acids are used in excess of methyl glycoside in order to increase reactivity, only monoesters can be selectively obtained with almost no by-products such as diesters, and therefore methyl glycoside fatty acids The present invention was based on the discovery that this method is industrially very advantageous as a method for producing monoesters.

The present invention will be explained in more detail below.

In the method for producing methyl glycoside fatty acid monoester of the present invention, the first raw material is a methyl glycoside of a sugar selected from monosaccharides having 5 to 7 carbon atoms and disaccharides consisting of hexoses, and hemiacetals (anomers) of the above sugars.
Those having a methyl group as an aglycone in the hemiacetal (anomeric) hydroxyl group are used, and the steric configuration after methyl substitution of the hemiacetal (anomeric) hydroxyl group is either α or β alone or a mixture of α and β in any proportion. Any of these can be used.

Here, the monosaccharide constituting the sugar moiety (glycone) is arabinose, which is a monosaccharide with 5 carbon atoms.

Examples include ribose, xylose, lyxose, xylulose, ribulose, 2-deoxyribose, etc. Monosaccharides with 6 carbon atoms include glucose, galactose, fructose, mannose, sorbose, talose, 2-deoxyglucose, 2-deoxygalactose, etc. Examples of monosaccharides with 7 carbon atoms include alloheptulose and sedoheptulose.

Examples include mannoheptulose and glucoheptulose. In addition, examples of disaccharides consisting of hexose include maltose, lactose, and the like.

In addition, in this invention, the said methyl glycoside can be used individually or in combination of 2 or more types.

Next, the second raw material of the production method of the present invention has 6 to 22 carbon atoms.
fatty acid or its lower alkyl ester.

Here, the fatty acid used in the present invention is a saturated or unsaturated linear or branched fatty acid having 6 to 22 carbon atoms, and such a fatty acid has a hydroxyl group, a carbonyl group,
It may also be substituted with a phenyl group or the like. in particular,
Fatty acids include caproic acid, sorbic acid, caprylic acid, capric acid, lauric acid, myristic acid, palmitoleic acid, palmitic acid, stearic acid, and isostearic acid.

Oleic acid, linoleic acid, linuric acid, pentadecanoic acid,
Eicosanoic acid, tocosanoic acid, tocosenoic acid.

Arachidonic acid, lysyllic acid, dihydroxystearic acid, etc. can be used.

Furthermore, as the fatty acid ester, the above-mentioned carbon number 6-22
It uses esters of fatty acids and lower alcohols having 1 to 3 carbon atoms, such as methanol, ethanol, and propatool, specifically methyl caproate, ethyl caproate, methyl caprate, ethyl caprate, and laurin. Methyl acid, prol laurate, propyl laurate, methyl myristate, ethyl myristate, propyl myristate, methyl palmitate, ethyl palmitate, propyl palmitate, methyl stearate, ethyl stearate, propyl stearate, methyl oleate , ethyl oleate.

Propyl oleate, methyl linoleate, ethyl linoleate, propyl linoleate, methyl linoleate, ethyl linoleate, propyl linoleate.

Examples include methyl eicosanoate, methyl arachidonate, methyl tocosanoate, and methyl tocosenoate.

In this case, the amounts of both of the above raw materials to be used are appropriately selected, but usually 0.05 to 0.05 to methyl glycoside per mole of fatty acids.
50 mol is used, preferably 0.1 to 10 mol. In the present invention, even if fatty acids are used in excess with respect to methyl glycoside, monoesters are preferentially obtained, and by-products of polysubstituted products such as diesters are kept extremely low.

In the present invention, both of the above raw materials are reacted in a specific organic solvent described below using a hydrolase, and the hydrolase used here is a neutral thermostable immobilized hydrolase. In this case, the hydrolytic enzymes include lipases such as porcine pancreatic lipase, yeast lipase derived from the genus Candida, bacterial lipase derived from the genus Aspergillus, Mucor, and Pseudomonas, esterase derived from porcine liver, trypsin, chymotrypsin, Examples include proteases such as subtilisin, but hydrolytic activity is limited to pH 5.5-8.
．． It has a maximum value in the range of 0 and is heat resistant.

And it is fixed.

For example, as a neutral heat-stable hydrolase, enzyme powder 50
■ 0.41d phosphate buffer (0.1M, pH 7)
) and the residual activity after heating at 70°C for 30 minutes is 40% or more, preferably 80% or more, more preferably 9
Various materials can be used as long as they have a heat resistance of 5% or more.
neutral thermostable lipase (sp-31S2. manufactured by NoVO, maximum activity pH 7,
5) Mucor Peng1ehei
)-derived neutral thermostable lipase (Lipozyme.

NoVO Co., Ltd., maximum active pH 6.0), etc. are preferably used. In addition, thermostable proteases include those derived from Bacillus thermobroteorixus (Thermolysin ■) and those derived from Thermus aquaticus YT-G (
Aqualysin 0) and the like are used, but of course they are not limited to these.

These neutral heat-stable hydrolytic enzymes may be purified products or crude products, and it is also possible to use dried bacterial cells (treated bacterial cells, resting or stationary bacterial cells) that produce hydrolytic enzymes.

Furthermore, as a method for immobilizing the neutral thermostable hydrolase, any of the carrier binding method, crosslinking method, and entrapping method may be employed, and the carrier binding method is particularly preferably employed.

In this case, specific immobilization carriers include activated carbon, porous glass, acid clay, bleaching clay, kaolinite, alumina, silica gel, bentonite, hydroxyapatite,
Inorganic substances such as calcium phosphate and metal oxides, natural polymer compounds such as starch and gluten, polyethylene and polypropylene.

Synthetic polymeric substances such as phenol-formalin resin, acrylic resin, anion exchange resin, and cation exchange resin can be mentioned, but in the present invention, synthetic polymeric substances that have porosity as a physical form, such as porous polyethylene, Porous polypropylene, porous phenol formalin resin, and porous acrylic resin are most preferably used. In the present invention, various immobilization carriers other than those described above may be used without any problem as long as they do not inhibit the expression of enzyme activity.

Furthermore, the amount of hydrolase immobilized on the immobilization carrier is usually 0.1 to 500 μg of protein per 1 g of the immobilization carrier, and in particular, the amount of hydrolase contained in the protein is about 2 to 50%. Preferably, the protein is immobilized.

In the present invention, the amount of the hydrolytic enzyme used is not particularly limited, but is 0.1 to 100 parts by weight of the fatty acids.
It can be in the range of 10,000 parts by weight, preferably 1 to 2,000 parts by weight.

In the present invention, the enzymatic reaction between methyl glycoside and the above-mentioned fatty acids using a neutral thermostable immobilized hydrolase is performed using a specific organic solvent that does not substantially contain water, that is, a dipolar moment of 0.5 Debye (Debye). ) in the presence of the above polar solvents. Examples of such organic solvents having a dipole moment of 0.5 debye or more are described in Table 11.172 on pages 1404 to 1406 of "Chemistry Handbook Basic Edition Revised 2nd Edition".

Specifically, secondary or tertiary alcohols are preferred;
For example, 2,4-dimethyl-3-pentanol, 2,6-
Cymethyl-4-heptatool, tertiary butyl alcohol, tertiary amyl alcohol.

Diacetone alcohol, 3-methyl-3-pentanol, 3-ethyl-3-pentanol, 3-propyl-3-
Pentanol, 2-methyl-2-hexanol, 2-ethyl-2-hexanol, etc. can be used. Furthermore, ketones such as acetone and methyl ethyl ketone, and dimethyl ether.

Ethers such as diethyl ether and dioxane, halogenated hydrocarbons such as chloroform and methylene dichloride, etc. are also suitably used, as well as pyridine.

Dimethylformamide, dimethylacetamide.

Nitrogen-containing solvents such as quinoline, sulfur-containing solvents such as dimethyl sulfoxide, etc. can also be used in combination.

Note that these solvents may be used alone or as a mixed solvent of two or more.

The amount of the organic solvent used depends on the type of organic solvent, the carbon chain length of the fatty acid or its ester, the reaction temperature, etc., but is preferably 10 to 99% by weight of the entire reaction system, especially 6% by weight of the entire reaction system.
It is 0 to 80% by weight.

When methyl glycoside and fatty acids are subjected to an enzymatic reaction using a hydrolase, the reaction conditions can be adjusted as appropriate, and the reaction proceeds even at low temperatures, but in order to accelerate the reaction rate, it is necessary to It is preferable to carry out the reaction at a certain temperature, and if the reaction is carried out at this temperature condition, the reaction can be completed in about 24 hours. In addition, even in such a high temperature reaction, there is no enzyme deactivation due to the use of a neutral heat-resistant immobilized hydrolase.

Furthermore, when producing methyl glycoside fatty acid monoester by the method of the present invention, for example, a method of filling a column with a hydrolase and passing a substrate solution (packed column method), a method of introducing a substrate solution and a hydrolase into a reaction tank, etc. The reaction can be carried out by employing a method in which the reaction is carried out by stirring or shaking (batch method), a method in which the reaction is carried out continuously in the batch method (continuous stirring tank method), or the like.

In this case, the method of the present invention is extremely advantageous industrially because the reaction can be carried out without deactivating the purified substance, so long-term continuous reactions and repeated batch reactions can be carried out without any problems.

In addition, in the method of the present invention, water or carbon number 1
~3 lower alcohol is produced as a by-product, but in this case, the concentration of this by-product in the system is 0.5% by weight or less, especially 0.1% by weight.
In order to proceed with the reaction efficiently, it is preferable to remove by-products as follows. Methods for removing these byproducts include, for example, zeolite, molecular sieves,
A method of adsorbing and removing Glauber's salt, etc. using outside and/or inside the reaction system, introducing dry air or inert gas into the reaction tank and removing it by evaporation into gas, or reducing the pressure inside the reaction tank. For example, the removal method may be evaporated, evaporated, and discharged out of the reaction tank.If these removal methods are appropriately combined with the above-mentioned enzyme reaction apparatus, the synthesis reaction can be carried out efficiently.

Note that the obtained reaction mixture can be purified according to a conventional method, and unreacted fatty acids contained in the reaction mixture can be separated and recovered for reuse.

The methyl glycoside fatty acid monoester thus obtained is an excellent surfactant and is used as an emulsifier in a wide range of fields such as foods, cosmetics, and pharmaceuticals.

〔Effect of the invention〕

According to the present invention, methyl glycoside fatty acid monoester can be efficiently obtained while suppressing the by-product of polysubstituted products such as diesters.

EXAMPLES Hereinafter, the present invention will be specifically explained with reference to Examples and Comparative Examples, but the present invention is not limited to the Examples below.

[Example 1] Methyl glucodide Ig (5.2 mmol), oleic acid 6 g (21.3 mmol), neutral heat-stable immobilized lipase [heat-stable lipase derived from Candeda antarctica immobilized on acrylic resin Shimo) (immobilized lipase 5
p-882, NoVO Co.)] 1100mg As a demethanol agent, 2g of Molecular Sieves 5A and 15ml of tertiary butyl alcohol were placed in a 50ml Erlenmeyer flask.
The mixture was shaken reciprocally at 60"C for 24 hours to carry out an esterification reaction.

After the reaction, the reaction solution was filtered to recover the enzyme, and the filtrate was concentrated using a rotary evaporator to remove the solvent. When this residue was separated by column chromatography, methyl-6-0-euler glucoside was obtained with a yield of 96%. Moreover, the production ratio of monoester and diester was 99.510.5.

[Example 2] 19.4 g (0.1 mol) of methyl glucodide and n-
93.0 g (0.5 mol) of methyl decanoate was added to 150 g of amyl alcohol, then 2 g of neutral heat-resistant immobilized lipase [Lipozyme (manufactured by NOVO)] was added, and 30 g of Molecular Sieves SA was added as a demethanol agent.
was added, and the mixture was reacted at 40"C for 24 hours.

After the reaction, the reaction solution was acetylated by a conventional method and analyzed by gas chromatography, which showed that methyl-6-0-decanoyl glucodide was obtained with a yield of 96% and a selectivity of 99.5%. confirmed.

[Example 3] 19.4 g (0.1 mol) of methyl glucodide and n-
50.5 g (0.3 mol) of decanoic acid was added to 120 g of diacetone alcohol, followed by addition of neutral thermostable immobilized lipase sp-382, and then at 40 °C under reduced pressure (3
0 mmHg) for 24 hours.

After the reaction, the reaction solution was acetylated by a conventional method and analyzed by gas chromatography, which showed that methyl-6-0-decanoyl glucodide was obtained in a yield of 97% and a selectivity of 99.3%. confirmed.

[Comparative example]

Example 1 except that 100 mg of alkaline pase PL-679 powder was used instead of the neutral heat-resistant immobilized lipase.
Esterification reaction was carried out in the same manner as above.

The reaction solution was operated in the same manner as in Example 1, and 20 ml of chloroform was added to the resulting residue to dissolve it, and then insoluble materials were removed by centrifugation. When this supernatant was separated by column chromatography, it was confirmed that methyl-6-0-oleyl glucodide was obtained, but the yield was 25
8 mg, and the yield was low (11%).

Applicant: Laion Co., Ltd.

Claims (1)

[Claims] 1. Methyl glycoside of a sugar selected from monosaccharides with 5 to 7 carbon atoms and disaccharides consisting of hexoses, and methyl glycosides with 6 to 7 carbon atoms.
22 saturated and unsaturated fatty acids and the fatty acids with 1 carbon number
A substantially water-free organic compound having electrical properties with a dipole moment of 0.5 debye or more is prepared by using a neutral heat-resistant immobilized hydrolase to convert fatty acids selected from esters with lower alcohols of A method for producing a methyl glycoside fatty acid monoester, which comprises reacting in a solvent.