JPH0670789A - Production of glyceroglycolipid - Google Patents

Abstract

PURPOSE:To easily obtain a glyceroglycolipid useful for low-caloric food, etc., by carrying out the condensation and transfer reaction of a fatty acid monoglyceride with a monosaccharide, polysaccharide or its derivative, etc., in a mixture of an organic solvent and an aqueous medium in the presence of a sugar hydrolyzing enzyme. CONSTITUTION:A fatty acid monoglyceride (e.g. monocapryl glycerol) and a monosaccharide (e.g. glucose) or di- or polysaccharide (e.g. lactose) or their derivatives are subjected to condensation and transfer reaction in a mixed solvent capable of dissolving the above compounds and consisting of an organic solvent (e.g. acetone) and an aqueous medium (e.g. buffering solution) in the presence of a sugar hydrolyzing enzyme originated from Escherichia coli (e.g. beta-galactosidase) at 20 deg.C for 48hr. The organic solvent is distilled out, the transfer reaction product is extracted with ethyl acetate, etc., and the obtained solid component is purified by a droplet counter-current partition chromatography to obtain the objective glyceroglycolipid useful as a raw material for low-caloric food, an emulsifier, a detergent, a humectant, a cosmetic base, etc.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a process for producing glyceroglycolipids useful as low calorie food materials, emulsifiers, detergents, moisturizers, cosmetic bases and the like.

[0002]

2. Description of the Related Art In recent years, researches have been actively conducted to enzymatically modify fats and oils to develop materials having new functions. Among them, many methods for modifying phospholipids with enzymes have been proposed, and some of them have been put into practical use. Even though glyceroglycolipid has the potential as a new material, it is presently present only in trace amounts in plants and has not been used so far.

Since glyceroglycolipid is a nonionic substance containing sugar as a hydrophilic group, it is less irritating to the skin when used as a detergent or cosmetic base, and p
There is little influence of H on the solubility. In addition, the high calorie content of fats and oils is currently a problem, and is expected as a low calorie food material. Further, in recent years, natural materials with excellent safety and biodegradability have been attracting attention from the standpoint of protecting the global environment.Glyceroglycolipids are natural surface-active substances contained in plants, and thus have high safety and biodegradability. Considered to be excellent.

Conventionally, as a method for producing glyceroglycolipid, an extraction method from a plant (Japanese Patent Laid-Open No. 178596/1987) has been used.
No.), a chemical synthesis method (JP-A-4-103594), an enzymatic synthesis method (European Patent No. 268461), etc., but all of them involve complicated steps, and particularly in the latter both synthesis methods, sugar and alcohol are used. It requires two or more steps of reacting and then reacting this with a fatty acid to synthesize,
Moreover, none of these are satisfactory in terms of productivity such as low yield, and an easy industrial production method of glyceroglycolipid has been desired.

[0005]

SUMMARY OF THE INVENTION It is an object of the present invention to provide a method capable of easily producing glyceroglycolipid in one step.

[0006]

Means for Solving the Problems As a result of intensive studies to achieve the above-mentioned objects, the present inventors have found that a fatty acid monoglyceride and a saccharide or a derivative thereof are dissolved in an organic solvent and an aqueous medium. We obtained the finding that glyceroglycolipids can be easily produced industrially by conducting a condensation / transfer reaction in the presence of a sugar hydrolase in a mixed solvent of 1), which enables high-yield synthesis in a one-step process. The present invention has been completed.

That is, in the method for producing a glyceroglycolipid of the present invention, a fatty acid monoglyceride and a monosaccharide or a disaccharide or higher saccharide or a derivative thereof are dissolved,
It is characterized in that the condensation / transfer reaction is carried out in the presence of a sugar hydrolase in a mixed solvent of an organic solvent and an aqueous medium. Hereinafter, the method of the present invention will be described in detail.

The glyceroglycolipid synthesized in the present invention is one in which one molecule of fatty acid monoglyceride and one molecule of monosaccharide are ether-bonded. Examples of monosaccharides constituting the glyceroglycolipid include xylose, glucose, fructose, mannose, galactose, and arabinose. As the fatty acid monoglyceride, glycerol and saturated or unsaturated linear or branched carbon atoms having 6 to 6 carbon atoms are used.
Consists of 24 fatty acid residues, such as caproic acid (C
_6), caprylic (C _8), capric acid (C _10), lauric acid (C _12), myristic acid (C _14), palmitic acid (C _16), palmitoleic acid (C _16: 1), stearic acid ( C ₁₈ ), oleic acid (C _{18: 1} ), linoleic acid (C _{18: 2} ), linolenic acid (C _{18: 3} ), arachidonic acid (C
_20: 4), eicosapentaenoic acid (C _20: 5), behenic acid (C _22), erucic acid (C _22: 1), docosahexaenoic acid (C _22: 6), tetracosahexaenoic tetra-enoic acid (C _{24: 4} ) And the like.

The reason for easily synthesizing the above glycolipids on an industrial scale is that, when an enzyme is used, the fatty acid monoglyceride as a substrate is poorly soluble in water, so that it is not efficient or does not react at all. Was never done industrially from. Therefore, in the present invention, by using a mixed solvent of an organic solvent and an aqueous medium, which dissolves both a fatty acid monoglyceride serving as a substrate and a saccharide of a monosaccharide or a disaccharide or a derivative thereof, the reaction is made efficient. In addition, it was possible to suppress hydrolysis, which is a side reaction, and to enable enzymatic synthesis of glyceroglycolipid.

That is, the present invention uses, as a reaction solvent, a mixed solvent of an organic solvent and an aqueous medium, which dissolves both a fatty acid monoglyceride serving as a substrate and a saccharide of monosaccharide or disaccharide or a derivative thereof. And it is necessary to dissolve the substrate as a whole of the mixed solvent. When the reaction is carried out only in an aqueous medium, as described above, the fatty acid monoglyceride serving as the substrate is poorly soluble, so that it is not efficient or does not react at all, on the other hand,
By adding an organic solvent, the solubility of the substrate is improved, and the equilibrium state of the reaction is changed, whereby hydrolysis, which is a side reaction, is suppressed, and the reaction proceeds efficiently.

The organic solvent used in the present invention is preferably one which is miscible with water, and examples thereof include acetone, acetonitrile, dioxane, pyridine, t-butanol, formamide, dimethylformamide and dimethylsulfoxide. These can be used alone or in admixture with a plurality of aqueous media, and can be used at such a ratio that both the fatty acid monoglyceride and the saccharides of monosaccharides or disaccharides or derivatives thereof are dissolved, but the ratio of the aqueous medium to the entire reaction system. Is preferably 5 to 60%. Water or various buffers are used as the aqueous medium.

The enzyme used in the reaction may be of any origin as long as it is a sugar hydrolase, but is preferably of microbial origin. For example, Escherichia coli (Escher
ichia coli) -derived galactosidase, yeast-derived glucosidase, yeast (Saccharomyces cerevisiae) -derived invertase, Aspergillus niger (Aspergillu)
s nigar) -derived xylosidase, etc., and galactosidase derived from E. coli is particularly preferable because it has a high transfer activity for fatty acid monoglyceride.

By immobilizing these enzymes on a suitable carrier, they can be stabilized against an organic solvent and can be used repeatedly. In the present invention, the saccharide serving as a substrate is a monosaccharide or a saccharide having a disaccharide or higher or a derivative thereof, and examples thereof include monosaccharides such as fructose, galactose, glucose, mannose, xylose and arabinose, phenylgalactopyranoside and nitrophenyl. Examples include monosaccharide derivatives such as glucopyranoside, disaccharides such as lactose, sucrose, maltose and xylobiose, sugar esters, oligosaccharides, agarose, carrageenan and pullulan.

The reaction temperature may be a temperature at which the enzyme is not completely deactivated, and is usually 5 to 70 ° C, preferably 10 to 50 ° C. The reaction pH is preferably set under optimum conditions for the enzyme used.

[0015]

EXAMPLES Next, the present invention will be specifically described by showing examples, but the present invention is not limited to the examples. (Example 1) [Synthesis of monocapryl monogalactosyl glycerol from lactose and monocapryl glycerol] 12 g of lactose was dissolved in 50 ml of 50 mM phosphate buffer (pH 7.3) containing 1 mM of magnesium chloride, and 20 ml of monocapryl glycerol was added. 50 ml of acetone was added and dissolved. To this, Escherichia coli
β-galactosidase (coli) (Wako Pure Chemical Industries) 1
000 units was added and the reaction was carried out at 20 ° C. for 48 hours. After the reaction, acetone was distilled off, and the transfer product was extracted with ethyl acetate. The product was isolated from the obtained solid by a droplet AC partition chromatography. The product released galactose by decomposition with galactosidase, and production of monocapryl monogalactosyl glycerol was confirmed by GC-MS analysis. The product was 15.7 mol% (yield).

(Example 2) [Synthesis of monofructomonolaurylglycerol from sucrose and monolaurylglycerol] Sucrose 1
2 g was dissolved in 50 ml of 25 mM acetate buffer (pH 5.0), and 20 g of monolaurylglycerol and 50 ml of t-butanol were added and dissolved. To this, 1000 units of yeast-derived invertase (manufactured by Sigma) was added to give 30
The reaction was carried out at ℃ for 24 hours. After the reaction, the product was separated by a silica gel column and isolated. The product was 10.5 mol% (yield).

(Example 3) [Synthesis of monocapryl monoglucosyl glycerol from glucose and monocapryl glycerol] 10 g of glucose was added to 50 ml of 50 mM phosphate buffer (pH 7.0).
20 ml of monocapryl glycerol and 50 ml of acetonitrile were added and dissolved. To this, 500 units of yeast-derived glucosidase (manufactured by Godo Shusei Co., Ltd.) was added and reacted at 30 ° C. for 24 hours. After the reaction, the product was isolated by separating with a silica gel column. The product was obtained in a yield of 13.5 mol%.

(Example 4) [Synthesis of monolauryl monogalactosyl glycerol from phenyl galactopyranoside and mono lauryl glycerol] 10 g of phenyl galactopyranoside was added to 50 mM phosphate buffer containing 1 mM of magnesium chloride (pH).
7.3) Dissolved in 50 ml, 20 ml of monolauryl glycerol and 50 ml of acetone were added and dissolved. In addition, β derived from Escherichia coli
-Galactosidase (manufactured by Wako Pure Chemical Industries, Ltd.) of 1000 units was added and reacted at 20 ° C for 24 hours. After the reaction, the product was extracted with ethyl acetate and then separated with a silica gel column to isolate the product. The product was obtained in a yield of 11.5 mol%.

[0019]

According to the present invention, a glyceroglycolipid can be easily produced in one step. The glyceroglycolipid obtained by the method of the present invention is suitable for use as a low-calorie food material, an emulsifier, a detergent, a moisturizer, and a cosmetic base.

Claims (1)

[Claims] 1. A condensation product of a fatty acid monoglyceride and a monosaccharide or a disaccharide or higher saccharide or a derivative thereof in a mixed solvent of an organic solvent and an aqueous medium in the presence of a sugar hydrolase. A method for producing a glyceroglycolipid, which comprises carrying out a transfer reaction.