JPS6314949B2 - - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method for producing fatty acid esters of sugars or sugar alcohols using enzymes.

Sugar fatty acid esters, typified by sucrose higher fatty acid esters, have conventionally been produced by a transesterification reaction in which sugar and fatty acid lower alkyl ester are reacted in the presence of an alkaline catalyst. Industrial production methods include a solvent method using dimethylformamide, which is a common solvent for sugar and fatty acid ester, and a method in which sugar is dissolved in propylene glycol or water and the fatty acid ester is made into a microemulsion in the presence of a fatty acid alkali metal salt. A microemulsion method in which sugar and fatty acid ester are reacted by dispersion, and a direct method in which sugar and fatty acid ester are melted together with a fatty acid alkali metal salt and reacted are known. Disadvantages of these methods include the unavoidable coloration of the product due to heating during the reaction process, and the use of dimethylformamide as a reaction solvent, which is unsuitable for the production of food additives. be.

In order to avoid these drawbacks, the present inventors have devised a method for producing sugar fatty acid esters from sugars and higher fatty acids using a hydrolytic enzyme such as lipase, as a method for carrying out the reaction at low temperatures and in an aqueous system.

The present invention uses an enzyme characterized in that trisaccharides, tetrasaccharides, polysaccharides, or sugar alcohols (excluding sorbitol and sorbitan) and higher fatty acids are incubated in the presence of a hydrolase having lipase activity. This invention relates to a method for producing sugar fatty acid esters.

Examples of sugar components and sugar alcohol components that can be used in the present invention include the following.

Trisaccharides: maltotriose, raffinose, cellotriose, manninotriose Tetrasaccharides: cellotetrose, stachyose Polysaccharides: dextrin, cyclodextrin, mannan, fructan, galactan, xylan,
Araban, cellulose, cellulose derivatives (CMC, hydroxypropylcellulose, methylcellulose) Sugar alcohols: arabitol, xylitol, mannitol, dulcitol, maltitol, lactitol Suitable higher fatty acids are saturated or unsaturated fatty acids with 8 to 22 carbon atoms. . Examples include:

Caprylic acid, capric acid, lauric acid, myristic acid, palmitic acid, stearic acid, arachic acid, behenic acid, caproleic acid, Linderic acid,
Myristoleic acid, palmitoleic acid, oleic acid, cadreic acid, erucic acid, decadienoic acid, linoleic acid, hiraric acid, linolenic acid, eicosatrienoic acid, docosatrienoic acid, hexadecatetraenoic acid, stearidonic acid, arachidonic acid, docosatetra Enoic acid, eicosapentaenoic acid, sardine acid,
Sabinic acid, iprolic acid, yarapinoleic acid, ricinoleic acid, feronic acid.

As is well known, hydrolytic enzymes having lipase activity include those derived from animals and those derived from microorganisms, and either of these may be used. For example, those derived from pig pancreas and those derived from microorganisms.
Aspergillus, Rhizopus, Pseudomonas,
Enterobacterium, Chromobacterium,
Some are derived from microorganisms such as Geotrichum, Penicillium, Mucor, and Candida genera. These enzymes do not necessarily need to be isolated and used; for example, crude enzymes such as pancreatin or commercially available enzyme preparations containing lipase can be used as they are.

Among them, those derived from Candida Cylindracea
Lipase MY (manufactured by Meito Sangyo Co., Ltd.) has a significantly high production yield of sugar or sugar alcohol ester.

The optimum pH for these enzymes is between 5 and 8;
A PH range of 4 to 9 may be used.

The reaction involves adding the substrate and enzyme to a buffer solution,
This is carried out by incubating at 20 to 60°C, preferably 30 to 50°C, until equilibrium is reached. The ratio of sugar or sugar alcohol to fatty acid is selected in the range of 6:1 to 1:6 (molar ratio), and the total substrate concentration is 1 to 30%, generally a few percent. Since fatty acids are sparingly soluble in buffer solutions, it is preferable to use them after finely pulverizing them, or emulsifying them with soap or the like that is harmless to enzymes. It is also preferable to stir constantly during the reaction. Compounds with low water solubility such as polysaccharides may be reacted using a solubilizer or a solvent.

The amount of enzyme added varies depending on the origin, type, potency, etc. of the enzyme, but it is sufficient as long as the reaction mixture contains a predetermined enzyme activity.

Since this reaction is reversible, equilibrium is reached after the reaction progresses to some extent. The reaction is stopped in this state, and the sugar or sugar alcohol fatty acid ester can be separated and purified from the reaction solution by a conventional method, and the unreacted fatty acid can be recovered.

The principles of the invention can be applied to enzymatic reactions using well-known immobilized enzymes attached to carriers by microencapsulation, matrixing, or covalent bonds. In this case, purification of the product is greatly facilitated, and it is also possible to carry out continuous reactions by flowing the substrate solution through a column packed with immobilized enzyme. Moreover, the enzyme used can be used repeatedly.

As described above, according to the present invention, coloring of the product is avoided because high-temperature heating is not required in the reaction process, it is safe because water is used as a medium, and free fatty acids are used as the raw fatty acid component, which is different from conventional methods. Compared to purely chemical transesterification methods, the present invention has significant advantages.

Examples of the present invention are shown below.

Example 1 2.00 g of commercially available lipase preparation (derived from Rhizopus),
3.64 g of mannitol and 11.3 g of oleic acid were added to 1000 ml of a phosphate buffer solution with a pH of 5.4, and the mixture was incubated at 40° C. for 72 hours while stirring with a magnetic stirrer.

The reaction mixture is freeze-dried, the resulting freeze-dried product is extracted with chloroform, and the extract is concentrated under reduced pressure.
The chloroform extract is dissolved in tetrahydrofuran, centrifuged at 3000 rpm, and separated into a tetrahydrofuran-soluble fraction and a tetrahydrofuran-soluble fraction.

Gel permeation chromatography was performed on the tetrahydrofuran soluble fraction, and the fraction eluting as the first peak was collected to obtain 6.60 g of mannitol oleate.

Example 2 10.08 g of raffinose, 11.30 g of oleic acid, and 4.0 g of a commercially available lipase preparation (derived from Candida) were placed in 1000 ml of a phosphate buffer solution of pH 5.4, and incubated at 40° C. for 72 hours while stirring with a magnetic stirrer. Thereafter, the same treatment as in Example 1 was carried out to obtain 6.65 g of ruffinose oleate.

Example 3 Cellotetrose 13.34g, oleic acid 11.3g,
4.0g of commercially available lipase preparation (derived from Candida) at PH5.4
Example 1 below.
By the same operation as above, 7.53 g of cellotetrose oleate was obtained.

Example 4 Carboxymethyl cellulose (degree of substitution 0.6) 10
g, 22.0 g of oleic acid, and 4.0 g of a commercially available lipase preparation (derived from Candida) in a phosphate buffer solution with a pH of 7.3.
The mixture was poured into a 1000 ml container and incubated at 40°C for 72 hours while stirring with a magnetic stirrer. Thereafter, the same treatment as in Example 1 was carried out to obtain 4.62 g of carboxymethylcellulose oleate.

Claims (1)

[Claims] 1. Sugar or sugar alcohol using an enzyme, which is characterized by incubating trisaccharides, tetrasaccharides, polysaccharides, or sugar alcohols (excluding sorbitol and sorbitan) and higher fatty acids in the presence of a hydrolase having lipase activity. A method for producing fatty acid esters.