JPH05148285A - Production of saccharides fatty acid ester - Google Patents

Abstract

PURPOSE:To maintain enzyme activity for a long period of time and to permit the repeated use of an expensive enzyme in reacting a saccharide with a fatty acid in the presence of a hydrolase in an organic solvent by adding a specific ion exchange resin to a reactional system. CONSTITUTION:In reacting (A) a saccharide selected from 5-7C monosaccharide, a disaccharide composed of hexose, 4-6C sugar alcohol, an ether compound composed of 5-7C monosaccharide and a monohydric alcohol and an ether compound composed of two kinds of hexoses and the monohydric alcohol with (B) a fatty acid selected from 6-22C fatty acid and an ester composed of the fatty acid and a lower alcohol by using a hydrolase in an organic solvent (preferably gamma-butyrolactone), an ion exchange resin selected from a styrenic chelate resin, a styrenic weakly basic anion exchange resin and an acrylic weakly basic anion exchange resin is added to the reactional system. Glucose, galactose, etc., are preferable as the component A.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to sugar fatty acid monoesters such as sugar fatty acid monoesters and alkylglycoside fatty acid monoesters (hereinafter, these are generically referred to simply as sugar fatty acid monoesters unless otherwise specified). The present invention relates to a method for producing a fatty acid ester of a saccharide that can be well synthesized and is therefore suitable for industrial production.

[0002]

2. Description of the Related Art Sugar fatty acid esters such as glucose fatty acid esters and alkyl glucoside fatty acid esters, especially monoesters, have excellent properties as surfactants, and therefore, the applicant has previously proposed that In addition to proposing blending sugar fatty acid monoesters with hair cosmetics and the like, various proposals have been made regarding methods for selectively producing sugar fatty acid monoesters (JP-A-3-765).
93, JP-A-4-16195, 16196).

These methods are methods of reacting a saccharide and a fatty acid with a hydrolase such as lipase in the presence of an organic solvent. The above method can selectively synthesize a sugar fatty acid monoester. In this respect, this method is superior to other conventional methods for producing sugar fatty acid ester.

Further, as a conventional method for producing a sugar fatty acid ester, the method of producing a sugar fatty acid ester by reacting a saccharide and a fatty acid with a hydrolase in the presence of an organic solvent has been used in the present application. In addition to the method by humans, JP-A-61-268192 and WO89 / 0
1480, WO90 / 09451, etc.

In these methods, in order to efficiently and advantageously obtain the fatty acid ester of the saccharide, it is desirable to repeatedly use an expensive enzyme, and thus it is desirable to maintain the activity of the enzyme for a longer period of time. To be done.

[0006]

Means and Actions for Solving the Problems The present inventors have
As a result of diligent study to meet the above demand, carbon number 5
~ 7 monosaccharides, disaccharides consisting of hexose, 4 carbon atoms ~
6 consisting of a sugar alcohol, an ether compound of a monosaccharide having 5 to 7 carbon atoms and a monohydric alcohol, and hexose 2
One or more types of saccharides selected from ether compounds of saccharides and monohydric alcohols, and one type of fatty acids selected from saturated and unsaturated fatty acids having 6 to 22 carbon atoms and esters of the fatty acids and lower alcohols Alternatively, when reacting two or more kinds with a hydrolase such as thermostable lipase in the presence of an organic solvent, a styrene chelate resin, a styrene weak basic anion exchange resin and an acrylic weak base are added to the reaction system. It was found that the enzyme activity can be maintained for a long period of time and the enzyme can be repeatedly used many times by adding one or more kinds of the cationic anion exchange resin, and thus the present invention can be made. I arrived.

The present invention will be described in more detail below. In the method for producing a fatty acid ester of sugar of the present invention, the first
The raw material is sugar. The sugar used in the present invention has 5 to 5 carbon atoms.
7 monosaccharides, disaccharides consisting of hexose, carbon number 4-6
1 or 2 or more kinds of saccharides selected from the above-mentioned sugar alcohol, an ether compound of a monosaccharide having 5 to 7 carbon atoms and a monohydric alcohol, and an ether compound of a disaccharide consisting of hexose and a monohydric alcohol.

Here, examples of monosaccharides having 5 carbon atoms include arabinose, ribose, xylose, lyxose, xylulose, ribulose and 2-deoxyribose.
As a monosaccharide having 6 carbon atoms, glucose, galactose, fructose, mannose, sorbose, talose, 2-
Deoxyglucose, 6-deoxygalactose, 6-
Examples thereof include deoxymannose and 2-deoxygalactose, and examples of monosaccharides having 7 carbon atoms include alloheptulose, sedoheptulose, mannoheptulose, and glucoheptulose.

Examples of the disaccharide composed of hexose include maltose, sucrose, sophorose and the like.

Further, examples of the sugar alcohol include erythritol, ribitol, xylitol, allitol, sorbitol, mannitol, galactitol and the like.

The ether compound of a monosaccharide having 5 to 7 carbon atoms or a disaccharide consisting of hexose and a monohydric alcohol is preferably a monohydric alcohol having 1 to 12 carbon atoms, particularly 1 to 6 carbon atoms. If you have alcohol like this,
The carbon chain may be linear, branched, saturated, unsaturated, unsubstituted or substituted, but preferably methyl alcohol, ethyl alcohol, propyl alcohol,
Butyl alcohol.

In this case, the ether bond position between the saccharide and the monohydric alcohol is not particularly limited and may be any position. Specific examples include glycosides such as methyl glucosid, ethyl glucosid, propyl glucosid, methyl fructoside, methyl mannoside, methyl maltoside, and methyl lactoside, 6-O-methyl glucose, 6-O-methyl fructose, and the like. The sugar ethers of As the alkyl glucosides, those having an alkyl group as an aglycone in the hemiacetal (anomeric) hydroxyl group of the saccharide are used, and the configuration after alkyl substitution of the hemiacetal (anomeric) hydroxyl group is α, β. Either of each of them or a mixture of α and β at an arbitrary ratio can be used.

Among the sugars mentioned above, glucose,
Fructose, galactose, mannose, maltose, sucrose, sorbitol, methyl glucosid, ethyl glucosid, propyl glucosid, butyl glucosid, methyl fructosid and methyl polyglucosid (maltosid, isomaltsid) are preferably used.

In the present invention, a monosaccharide having 5 to 7 carbon atoms having no substituent, a disaccharide consisting of hexose,
In addition, one or more kinds of saccharides selected from sugar alcohols having 4 to 6 carbon atoms and one or more kinds of ether compounds of the above-mentioned sugars and monohydric alcohols can be mixed and used together, depending on the combination ratio. A mixture of sugar fatty acid ester and sugar ether fatty acid ester can be synthesized all at once in a ratio and with high production efficiency.

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

Here, the fatty acid used in the present invention is a saturated or unsaturated, straight-chain or branched-chain fatty acid having 6 to 22 carbon atoms, and if it is such a fatty acid, a hydroxyl group, a carbonyl group, a phenyl group, etc. It may be replaced with.
Specifically, as fatty acids, caproic acid, sorbic acid, caprylic acid, capric acid, lauric acid, myristic acid, palmitoleic acid, palmitic acid, stearic acid, isostearic acid, oleic acid, linoleic acid, linolenic acid, pentadecanoic acid, eicosane Acids, docosanoic acid, docosenoic acid, arachidonic acid, ricinoleic acid, dihydroxystearic acid and the like can be used.

Further, as the ester of fatty acid, an ester of the above-mentioned fatty acid having 6 to 22 carbon atoms and a lower alcohol such as methanol, ethanol or propanol is used, and specifically, methyl caproate or ethyl caproate is used. , Methyl caprate, ethyl caprate, methyl laurate, ethyl laurate, propyl laurate, methyl myristate, ethyl myristate, propyl myristate, methyl palmitate, ethyl palmitate, propyl palmitate, methyl stearate, stearin Ethyl acid salt, propyl stearate, methyl oleate, ethyl oleate, propyl oleate, methyl linoleate, ethyl linoleate, propyl linoleate, methyl linolenate, ethyl linolenate, propyl linolenate, eicosane Methyl, methyl arachidonate,
Examples include methyl docosanoate and methyl docosenoate.

In this case, the amounts of both of the above raw materials used are appropriately selected. Usually, fatty acids of 0.9 to 2 per mol of sugar are used.
0 mol is used, preferably 1 to 10 mol, more preferably 2 to 5 mol. In this case, if the molar ratio of fatty acids is increased, the reaction rate increases, but the reaction rate does not increase even if it is used in excess of 20 mol. Therefore, from the economical point of view, the amount of fatty acids used is 20 mol or less. It is preferable that In the present invention, even if fatty acids are used in excess with respect to sugars, monoesters are preferentially obtained, and by-products of polysubstituted products such as diesters are suppressed to a very low level.

In the present invention, both of the above raw materials are reacted with a hydrolase in a specific organic solvent described later. Examples of hydrolases used here include porcine pancreatic lipase, yeast lipase derived from Candida, lipases such as bacterial lipase derived from Aspergillus, Mucor, and Pseudomonas, esterase derived from pig liver, trypsin, chymotrypsin. , And proteases such as subtilisin. Of course, it may be a lipase or the like produced by introducing these yeast DNAs into a host and producing the host. Among these, it has heat resistance,
The hydrolysis activity is pH 5 to 10, more preferably 5.
Those having a maximum value in the range of 5 to 9.5 are preferable.

For example, the thermostable hydrolase is an enzyme
50 mg of powder was added to 0.4 ml of phosphate buffer (0.1
M, pH 7) and after heating at 70 ° C for 30 minutes
Residual activity is 40% or more, preferably 80% or more, more preferably
Various if it has heat resistance of 95% or more
Can be used, and is disclosed in Japanese Patent Publication No. 1-501120.
Those described in the lipases listed above are preferably used.
It Specifically, Candida Antarctica (C
anda antarctica) derived heat resistance
Pase, Candida Tsukubaensis (Candid
atsukubaensis, ATCC 2455
5) Heat-resistant lipase derived from Candida auricla
Riae (Candida auriculariae,
ATCC 24121) derived thermostable lipase, can
Dida Fumicola (Candidahumicol)
a, ATCC 14438) -derived thermostable lipase,
Candida forlium
arum, ATCC 18820) derived heat resistant lipers
Ze, Mucor Miehei
Examples thereof include heat-resistant lipase derived from the above. Well
As a thermostable protease, Bacillus thermob
Derived from Roteolyxus (thermolysin , Trademark),
Derived from Thermus aquaticus YT-G (Aqua
Lysin , Trademark), etc. are used, but of course they are not limited to these.
It is not something that can be done.

The hydrolase may be a purified product or a crude product, and it is also possible to use a dried product of cells that produce the hydrolase (treated cells, resting or resting cells).

The above-mentioned hydrolase can be used after being immobilized, and as the immobilizing method, any of the carrier binding method, the cross-linking method and the entrapping method may be adopted. In particular, the carrier binding method can be preferably adopted.

In this case, specifically as the immobilization carrier,
Activated carbon, porous glass, acid clay, bleaching clay, kaolinite, alumina, silica gel, bentonite, inorganic substances such as hydroxyapatite, calcium phosphate, metal oxides, natural polymer compounds such as starch and gluten, polyethylene, polypropylene, phenol formalin resin,
Synthetic polymer substances such as acrylic resin, anion exchange resin, cation exchange resin, etc. can be mentioned, but in the present invention, synthetic polymer substances having a physical form of porosity, for example, 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 the above may be used as long as they do not inhibit the activity expression of the enzyme.

Further, the amount of hydrolase immobilized on the immobilizing carrier is usually 0.1 to 50 per 1 g of the immobilizing carrier.
0 mg of protein, especially hydrolase 2 to protein
It is preferable to immobilize a protein containing about 50%.

The amount of the hydrolase used in the present invention is not particularly limited, but is preferably 0.02 to 1 part by weight, more preferably 0.05 to 1 part by weight with respect to 1 part by weight of the saccharide.
0.8 parts by weight, more preferably 0.08 to 0.6 parts by weight. If the amount of the enzyme is too small, the reaction rate tends to be slow, while if the amount of the enzyme is too large, the by-product rate of the polyester of diester or more tends to increase.

Various kinds of organic solvents can be selected for use in the present invention. Solvents listed in JP-A-3-76593 and the like, for example, tertiary alcohols, heptanone, pentanone, ethylpyridine, pyridine, picolines, etc. In particular, the applicant of the present invention has previously filed Japanese Patent Application No. 3-1004.
It is preferable to use one kind or two kinds or more selected from acetylacetone, ethylene carbonate, propylene carbonate and γ-butyrolactone proposed in No. 84.

That is, when acetylacetone, ethylene carbonate, propylene carbonate or γ-butyrolactone is used as the organic solvent, the conversion rate is high and the monoester production rate is high, and the reaction between the saccharide and the fatty acid is converted within about 10 hours. The reaction can be completed at a rate of 90% or more, and the reaction is completed without increasing the amount of diester produced even for about 2 hours by increasing the enzyme concentration, which is advantageous for factory production.

Further, in this case, the amount of these solvents used is such that the reaction proceeds at least satisfactorily and therefore the reaction can be carried out with an increased substrate concentration, so that the reaction tank required to react the same amount of substrate is used. The volume of the solvent can be reduced, the cost of the production apparatus can be reduced, and unlike the solvent such as diacetone alcohol, since the above solvent does not cause thermal decomposition, the recovery rate from the reaction system is also after the reaction is completed. It is high and can be repeatedly recovered even in the reaction at 70 ° C. or higher. Further, when water or lower alcohol which is generated as a side reaction in the reaction system is removed during the reaction, the above solvent is used even if a simple decompression method is adopted. Since it is not accompanied by water or lower alcohol, only water or lower alcohol can be substantially removed and recovered, which is also advantageous from the viewpoint of factory production.

The amount of the above-mentioned organic solvent used depends on the type of the organic solvent, the carbon chain length of the fatty acid or its ester, the reaction temperature, etc., but is preferably 1 to 10 times by weight, more preferably 2 to 1 times the weight of the saccharide. 6 times the weight. In this case, it is possible to use a large amount of the above organic solvent, but in terms of reaction rate and particularly downsizing the reactor, to reduce the construction cost, reduce the running cost with the downsizing of the reactor. From the viewpoint of avoiding an increase in cost burden in factory production, it is recommended to use 10 times or less the weight of sugar as described above. In particular, by using acetylacetone, ethylene carbonate, propylene carbonate, and γ-butyrolactone, the solvent amount at least suppresses the by-product of the polyester such as diester and preferentially and efficiently converts the monoester into a high conversion rate and reaction rate. Monoesters can be produced.

In the present invention, one or two of styrene-based chelate resin, styrene-based weakly basic anion exchange resin, and acrylic-based weakly basic anion exchange resin are further used in the reaction system.
Add more than one seed.

In this case, as these resins, those having the structures of the following formulas (1) to (5) are preferable.

[0032]

[Chemical 1] (In the formula, M is a hydrogen atom or a metal atom such as Na, and n is an integer of 1 or more.)

As the above resin, commercially available products, DIAION CR-10, CR-20, WA- manufactured by Mitsubishi Kasei Co., Ltd.
20, WA-21, WA-10, WA-11, WA-3
0, Amberlite IRC-718 manufactured by Organo Corporation,
IRA-68, IRA-35, IRA-93ZU, IR
A-94S, Amberlyst A-21, etc. can be specifically used.

The amount of the above resin added is preferably 0.1 to 3 times by weight, more preferably 0.5 to 2 times by weight, the amount of the immobilized enzyme. If it is less than 0.1 times by weight, the effect of adding the resin may not be exhibited, and if it is more than 3 times by weight, the reaction rate may be lowered.

When enzymatically reacting saccharides and fatty acids with a hydrolase, the reaction conditions can be adjusted appropriately, and the reaction proceeds even at low temperatures, but in order to accelerate the reaction rate, it is 40 ° C. or higher, particularly 50 to 100. ℃, more preferably 60 ~ 90 ℃
It is preferable to react at a temperature of, especially as a reaction solvent acetylacetone, ethylene carbonate, propylene carbonate,
When γ-butyrolactone is used, if the reaction is performed under the above temperature conditions, the conversion rate is usually 90% in a short time of 2 to 10 hours.
The reaction can be completed as described above. Even in such a high temperature reaction, the enzyme is not deactivated by using the thermostable hydrolase.

In this case, according to the present invention, the monoester in the total amount of fatty acid ester of the synthesized saccharide is 95 to 100.
%, Diester is 0 to 5%, and the polyester of tri or more is the balance, particularly 97 to 100% of monoester, 0 to 3% of diester, and the polyester is the balance of tri or more.

Further, when the sugar fatty acid monoester is produced by the method of the present invention, for example, a method in which a substrate solution and a hydrolase are introduced into a reaction tank and the reaction is carried out by stirring and shaking (batch method), the batch method is used. It can be carried out by employing a method of continuously carrying out the reaction (continuous stirring tank type) or the like.

In this case, in the present invention, by adding the above resin, the activity of the enzyme can be maintained for a long period of time and the reaction can be carried out without deactivating the enzyme. Therefore, long-time continuous reaction or repeated batch reaction is possible. Since it can be carried out without trouble, it is extremely advantageous industrially. For example, when acetylacetone, ethylene carbonate, propylene carbonate, or γ-butyrolactone is used as a solvent, the effect can be effectively retained and the enzyme can be repeatedly used many times.

In the method of the present invention, water or a lower alcohol is by-produced by the enzymatic reaction. In this case, the concentration of this by-product in the system is 0.5% by weight or less, particularly 0.1% by weight or less. It is preferable to remove the by-product in order to efficiently proceed the reaction. As a method of removing these by-products, for example, a method of adsorbing and removing zeolite, molecular sieves, mirabilite, etc. outside and / or inside the reaction system, and introducing dry air or an inert gas into the reaction tank. Examples of the method include removing by evaporating in gas, or reducing the pressure in the reaction tank and evaporating and discharging it out of the reaction tank. It can be performed.

In this case, especially when acetylacetone, ethylene carbonate, propylene carbonate, γ-butyrolactone or the like is used as the reaction solvent, the boiling point thereof is remarkably different from that of water or lower alcohol which is by-produced as the reaction progresses. In addition, the gas introduction method and the pressure reduction method can be adopted without any trouble, because the apparatus is simple and has a low cost load.For example, by attaching a reflux condenser for the purpose of proper partial condensation, there is no accompanying reaction solvent and no by-product is produced. It is possible to collect only incoming water and lower alcohols in high concentrations. Therefore, it becomes easy to reuse the lower alcohol thus recovered, and also from this point, the manufacturing cost can be reduced.

When a dephlegmator is used, the degree of vacuum during the reaction, the temperature of the refrigerant of the dephlegmator and the temperature of the refrigerant of the cold trap are by-produced with the vapor pressure of the reaction solvent at the reaction temperature and the progress of the reaction. It is selected in consideration of the vapor pressure of incoming water or lower alcohol, but from the viewpoint of the reaction rate, as long as it is possible to reflux only the reaction solvent by appropriately adjusting the temperature of the refrigerant, the higher the degree of vacuum during the reaction is. desirable. The degree of vacuum is appropriately selected depending on the type of reaction solvent and other reaction conditions, and is practically 200 torr or less. For example, when γ-butyrolactone is used as the reaction solvent and methyl caprylate is used as the fatty acids and the reaction is carried out at 80 ° C., preferably 20
It is not more than torr, more preferably not more than 15 torr, still more preferably not more than 12 torr.

The obtained reaction mixture can be purified by a conventional method, and unreacted fatty acids contained in the reaction mixture can be separated, recovered and reused.

The sugar fatty acid monoester thus obtained (note that monosaccharides having 5 to 7 carbon atoms, disaccharides composed of hexose, and sugar alcohols having 4 to 6 carbon atoms are used as raw materials for these sugars). A fatty acid monoester is obtained, which is an ether compound of a monosaccharide having 5 to 7 carbon atoms and a monohydric alcohol,
Alkyl glycoside fatty acid monoesters are obtained when an ether compound of a disaccharide consisting of hexose and a monohydric alcohol is used as a raw material), all of which have excellent surface-active ability and are effectively used in various fields. ..

[0044]

EXAMPLES The present invention will be specifically described below with reference to examples, but the present invention is not limited to the following examples.

Examples 1 to 5 Methyl glucosid, methyl caprylate, γ-butyrolactone and Candida antarctica were placed in a four-neck round bottom flask equipped with a stirrer, thermometer, mercury manometer and reflux condenser. A neutral heat-resistant lipase derived from immobilization on an acrylic resin (immobilized lipase, manufactured by NOVO) was charged in the amount shown below, and the resin shown in Table 1 was added in the amount shown in the same table, and the above-mentioned reflux was performed. A dry ice / acetone trap and a water pump were connected to the outlet of the cooler, and the reaction was carried out at 80 ° C. under reduced pressure. In this case, the cooling water temperature of the reflux condenser is such that only the reaction solvent, γ-butyrolactone, is refluxed, and methanol, which is a byproduct of the reaction, and a small amount of water contained in the raw material are collected by the trap, so that the degree of vacuum is set to Considering this, the temperature was set to 30 ° C. Fatty acids / sugar = 4 (molar ratio) Reaction solvent / sugar = 4 (weight ratio) Enzyme / sugar = 0.1 (weight ratio) Reaction temperature 80 ° C., reflux under reduced pressure

After reacting for a predetermined time, the pressure was returned to normal pressure, stirring was stopped, and the mixture was allowed to stand in a room temperature atmosphere for 1 hour to precipitate the immobilized enzyme. Then, only the supernatant reaction solution was carefully collected with a pipette to a depth just before the immobilized enzyme was mixed in the pipet.

Next, raw materials other than the enzyme were newly charged into a four-necked round bottom flask in which the immobilized enzyme and a small amount of the reaction solution remained so that the above-mentioned conditions were newly established.
A second reaction was performed.

Thereafter, the enzyme was repeatedly used in the same manner as above, and the reaction was performed a predetermined number of times.

Table 1 shows the results of the reaction time, conversion and diester content in the above repeated reaction. The conversion rate and diester content were determined by sampling the reaction mixture at a suitable time, acetylating the product by a conventional method, and analyzing by gas chromatography to determine the amount of methyl glucoside caprylic acid ester (monoester and diester) and conversion. The rate was calculated.

[0050]

[Table 1] (Note): The diester content was calculated from the following formula.

[0051]

[Equation 1]

[0052]

EFFECTS OF THE INVENTION According to the present invention, when a saccharide fatty acid ester is produced by reacting a saccharide and a fatty acid with a hydrolase in the presence of an organic solvent, the activity of the enzyme is maintained for a long period of time. However, expensive enzymes can be used repeatedly.

Claims (2)

[Claims] 1. A monosaccharide having 5 to 7 carbon atoms, a disaccharide composed of hexose, a sugar alcohol having 4 to 6 carbon atoms, and 5 carbon atoms.
One or more saccharides selected from ether compounds of monosaccharides of 7 and monohydric alcohols and ether compounds of disaccharides of hexoses and monohydric alcohols, and saturated and unsaturated carbon atoms of 6 to 22 1 of fatty acids selected from fatty acids and esters of the fatty acids with lower alcohols
In the method for producing a fatty acid ester of a saccharide, which comprises reacting one or more species with a hydrolase in the presence of an organic solvent, a styrene chelate resin, a styrene weakly basic anion are added to the reaction system. A method for producing a fatty acid ester of a saccharide, comprising adding one or more of an exchange resin and an acrylic weakly basic anion exchange resin. 2. The method for producing a fatty acid ester of a saccharide according to claim 1, wherein one kind or two or more kinds selected from acetylacetone, ethylene carbonate, propylene carbonate and γ-butyrolactone is used as the organic solvent.