JPH0416196A - Production of alkylglycoside monoester of fatty acid - Google Patents

Abstract

PURPOSE:To enable efficient obtaining of the subject monoester by reacting alkylglycosides of sugars selected from disaccharides, etc., and specific fatty acids with a neutral thermoduric immobilized hydrolase in an organic solvent. CONSTITUTION:(A) Alkylglycosides of sugars having 1-6C alkyl groups as an aglycone on hemiacetal (anomeric) hydroxyl groups of the sugars are obtained by selection from 5-7C monosacharides, such as ribose as disaccharides composed of hexoses. (B) Fatty acids such as caproic acid (methyl caproate) 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) A neutral thermoduric immobilized hydrolase is subsequently obtained by immobilizing a porcine pancreatic lipase, etc., on active carbon, etc. To 1mol ingredient (B), is added 0.05-50mol ingredient (A). The ingredient (C) is then added so as to afford 0.1-10000 pts.wt. amount of the ingredient (C), based on 100 pts.wt. ingredient (B). Reaction is subsequently carried out in a solvent, such as tertiary butanol without containing water, at 50-120 deg.C for about 24hr, to produce the objective alkylglycoside of the fatty acids.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention is directed to the production of alkyl glycoside fatty acid esters from fatty acids and alkyl glycosides having an alkyl group having 1 to 6 carbon atoms by enzymatic reaction. The present invention relates to a method for producing an alkyl glycoside fatty acid monoester that can be synthesized.

Conventionally, as a method for producing alkyl glycoside fatty acid esters, a chemical synthesis method has been proposed in which a hexasaccharide 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 Box 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 C4 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 such as those whose alkyl groups are methyl or ethyl is ineffective, and in the latter method, the use of lower alkyl glycosides, such as methyl or ethyl, has the disadvantage of being ineffective. It is difficult to obtain alkyl glycoside fatty acid esters with monoester content. In particular, to increase reaction efficiency,
Increasing the amount of fatty acids relative to alkyl glycosides
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. An object of the present invention is to provide a method for producing a Cel C6 alkyl glycoside fatty acid monoester.

In order to achieve the above object, the present inventors conducted intensive studies and found that alkyl glycosides of sugars selected from monosaccharides having 5 to 7 carbon atoms and disaccharides consisting of hexoses (provided that The number of carbon atoms in the group is 1 to 6), and the number of carbon atoms in the group is 6
When reacting fatty acids selected from ~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 hydrolyzed abrasive is used, and tertiary When the reaction is carried out in a substantially water-free organic solvent selected from butanol, tertiary amyl alcohol, diacetone alcohol, chloroform, acetone, tetrahydrofuran and 1°4-dioxane, the alkyl group becomes a methyl group. Ester synthesis or transesterification reactions with the above-mentioned fatty acids can be carried out easily, efficiently, and favorably for lower alkyl glycosides such as ethyl or ethyl groups, and alkyl glycosides with a larger number of carbon atoms, and can be applied over a wide range of alkyl groups. Esterification by enzymatic reaction is possible, and not only a high reaction rate is achieved, but the selectivity of the reaction is also high, and there is almost no by-product of polysubstituted products such as diesters (diacyls),
It was found that only the monoacyl form was preferentially given.

In this case, even if fatty acids are used in excess of the alkyl glycoside in order to increase reactivity, only monoesters can be selectively obtained with almost no by-products such as diesters. The present invention was based on the discovery that this method is industrially very advantageous as a method for producing alkyl glycoside fatty acid monoesters.

The present invention will be explained in more detail below.

In the method for producing an alkyl glycoside fatty acid monoester of the present invention, the first raw material is an alkyl glycoside of a saccharide selected from disaccharides consisting of monosaccharides having 5 to 7 carbon atoms and hexoses, and hemiacetals (anomers) of the above saccharides. A hemiacetal (anomer) having an alkyl group of C to C6 as an aglycone in the hydroxyl group is used, and the steric configuration after alkyl substitution of the hemiacetal (anomer) hydroxyl group is α and β each alone, or α and β are in an arbitrary ratio. Any of the mixtures can be used. In addition, as long as the alkyl group has 1 to 6 carbon atoms, it may be linear, branched, saturated, unsaturated, unsubstituted, or substituted, but
Preferred are methyl, ethyl, propyl, and butyl groups.

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 alkyl 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, wasillic 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, ethyl 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 alkyl glycoside per mole of fatty acid.
~50 mol is used, preferably 0.1-10 mol. In the present invention, even if fatty acids are used in excess with respect to the alkyl 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, subtilisin, etc. proteases, etc., but the hydrolytic activity is at 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
■ 0.4all phosphate buffer (0, IM, pH
7) Various materials can be used as long as they have a heat resistance of 40% or more, preferably 80% or more, and more preferably 95% or more of residual activity after being dissolved in and heated at 70'C7 for 30 minutes. , Candida antarctica (C
andida antarctica)-derived neutral thermostable lipase (sp-382, manufactured by NoVO, maximum activity P
H7,5), Mucormie
hei)-derived neutral thermostable lipase (Lipozym
e.

NoVO Co., Ltd., maximum active pH 6.0), etc. are preferably used. Also, as a thermostable protease, Bacillus? Those derived from Thermobroteorixus (thermolysin■), those derived from Thermus aquaticus YT-G (
Aqualysin (■) 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 an alkyl glycoside and the above-mentioned fatty acids using a neutral heat-stable immobilized hydrolase is carried out using a specific organic solvent substantially free of water, such as tertiary butanol, tertiary amyl alcohol, etc. .

Diacetone alcohol, chloroform, acetone.

The reaction is carried out in the presence of one or more organic solvents selected from tetrahydrofuran and 1,4-dioxane.

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 alkyl glycosides 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 an alkyl 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 enzyme, and long-term continuous reactions or repeated batch reactions can be carried out without any problems.

In addition, in the method of the present invention, water or
~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, recovered, and reused.

The thus obtained C-CG alkyl glycoside fatty acid monoester is an excellent surfactant, and is suitable for foods,
It is used as a milk north side in a wide range of fields such as cosmetics and pharmaceuticals.

〔Effect of the invention〕

According to the present invention, C--CG alkyl glycoside fatty acid monoester can be efficiently obtained while suppressing the by-production 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] 1.04 g (5 mmol) of ethyl glucodide and 1.8 g (1 mmol) of methyl octoate were added to 100 ml of tertiary butyl alcohol to prepare a neutral heat-resistant immobilized lipase [derived from Candeda antarctica]. heat-resistant lipase immobilized on acrylic resin (immobilized lipase sp
-382, manufactured by NoVO)) using 0.5 g, 60
The mixture was stirred at ℃ for 24 hours to carry out the transesterification reaction.

After the reaction, the enzyme was removed, the solvent was distilled off under reduced pressure, and the residue was separated by column chromatography, resulting in ethyl-6-
0-octanoylglucodide was obtained with a yield of 98%. Also, the production ratio of monoester and diester is 9
It was 9.710.3.

[Comparative example]

1.04 g (5 mmol) of ethyl glucodide, 1.8 g (l1 mmol) of methyl octoate and 1 mol of 2-butanone.
In addition to 00ml, neutral fixed lipase [RibozyA (
(manufactured by NOVO)) was stirred at 60° C. for 24 hours to perform a transesterification reaction.

Then, when the same operation as in Example 1 was carried out, ethyl-6
Although it was confirmed that -0-octanoylglucodide was obtained, the yield was 10%, which was low.

[Examples 2 to 7] Various alkyl glycoside fatty acid monoesters were produced under the raw material conditions shown in the table below.

Claims (1)

[Claims] 1. An alkyl glycoside of a saccharide selected from monosaccharides having 5 to 7 carbon atoms and disaccharides consisting of hexoses (provided that the alkyl group has 1 to 6 carbon atoms) and 6 to 22 carbon atoms.
saturated and unsaturated fatty acids, and fatty acids with carbon atoms of 1 to 3
tertiary butanol, tertiary amyl alcohol, diacetone alcohol, chloroform, acetone, tetrahydrofuran and 1,4 - A method for producing an alkyl glycoside fatty acid monoester, which comprises reacting in a substantially water-free organic solvent selected from dioxane.