JPS60227687A - Production of sugar alcohol fatty acid esters - Google Patents

Abstract

PURPOSE:The incubation of sorbitol or sorbitan and a higher fatty acid is effected in the presence of a hydrase to produce a sugar alcohol fatty acid esters which are suitable for use as food additives. CONSTITUTION:Lipase is used as a hydrase. Any lipase, e.g., originating from animals or microorganisms can be used, however, the lipase from Candida cylindracea shows extremely high yield of sugar alcohol ester. Sorbitol or sorbitan and a higher fatty acid are mixed at a weight ratio of 6/1-1/6 to form an aqueus solution containing the total substrates in 1-30wt% concentration. The solution is adjusted in pH to 5-8, lipase is added to effect incubation at 20- 60 deg.C to form a sugar alcohol fatty acid ester. Since the reaction is reversible, it is stopped at the time the equilibrium is reached.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method for producing sugar alcohol fatty acid esters 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 dispersed as a microemulsion in the presence of a fatty acid alkali metal salt. A microemulsion method in which a sugar and a fatty acid ester are reacted by melting them together with a fatty acid alkali metal salt, and a direct method are known. The disadvantages of these methods are that coloration of the product due to heating during the reaction process can be avoided;
Furthermore, when dimethylformamide is used as a reaction solvent, it is unsuitable for producing food additives.

In order to avoid these drawbacks, the present inventors first discovered a method for producing sugar fatty acid esters from sugars and higher fatty acids using hydrolytic enzymes such as lipase, as a method of conducting the reaction at low temperatures and in an aqueous system. Ta. We discovered that this could be further applied to sugar alcohols, leading to our work.

The present invention resides in a method for producing a sugar alcohol fatty acid ester using an enzyme, which is characterized by incubating sorbitol, toll, or sorbitan and a higher fatty acid in the presence of a hydrolase.

Sugar alcohol components that can be used in the present invention include sorbitol and sorbitan.

Suitable higher fatty acids are saturated or unsaturated fatty acids having 8 to 22 carbon atoms.

As the enzyme, a hydrolytic enzyme, especially a lipase, is used. As is well known, there are lipases of animal origin and those of microbial origin; either of these may be used. For example, Asper is derived from pig pancreas, and Asper is derived from microorganisms.
gillus, Rh1zopus, Pseu-do
monas, Enterobacterium, C
hromobacterium.

Geotrichum, Penicillium,
Some are derived from microorganisms such as Mucor and Candida. These enzymes do not necessarily need to be isolated and used; for example, as crude enzymes such as vancreatin,
Alternatively, a commercially available enzyme preparation containing lipase can be used as is.

The optimum pH for these enzymes is between 5 and 8, although a pH range of between 4 and 9 can be used.

The reaction is carried out by adding the substrate and enzyme to a buffer solution and incubating at 20-60°C, preferably 30-50°C, until equilibrium is reached. The ratio of sugar alcohol to fatty acid is 6:1 to 1:
6 (weight ratio), and the total substrate concentration is 1 to 3.
0%, generally several % is used. 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.

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. In this state, the reaction is stopped, and the sugar alcohol fatty acid ester is purified from the reaction solution by a conventional method to recover unreacted fatty acids.

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 Commercially available lipase preparation (derived from Candida) 2', OOg
.

Add 3.64 g of sorbitol and 22.56 g of oleic acid to 1000 d of phosphate buffer at pH 5.4, stir with a magnetic cooler, and heat to 72 ml at 40 ''C.
Incubated for hours.

The reaction mixture is freeze-dried, the resulting freeze-dried product is extracted with chloroform, and the extract is concentrated under reduced pressure.

Dissolve the chloroform extract in tetrahydrofuran and add 30
The bones were centrifuged at 00 r, p, m and separated into tetrahydrofuran soluble and tetrahydrofuran insoluble components.

Gel permeation chromatography was performed on the tetrahydrofuran soluble fraction, and the fraction eluting as the first peak was collected, and sorbitol oleate 7,
49 g was obtained.

Example 2 In Example 1, 22.56 g of oleic acid (tL, t
), except that 22.8 g of stearic acid was used, and sorbitol stearate 6°27
I got g.

Example 3 Sorbitol 5.46g, oleic acid 22.56g.

4.0 g of commercially available lipase preparation (derived from Candida) was added to p
The mixture was placed in H7,3 phosphate buffer solution 10007d and incubated at 40° C. for 72 hours while stirring with a magsotic cooler. Thereafter, the same treatment as in Example 1 was carried out to obtain 16.92 g of sorbitol oleate.

Example 4 Sorbitan 3.28g, oleic acid 22.56g.

2.0 g of commercially available lipase preparation (derived from Candida) was added to p
The mixture was placed in a 1,000 volume H7,13 phosphate buffer solution, and the same procedure as in Example 1 was performed to obtain 8.65 g of sorbitan oleate.

Example 5 Sorbitan 4.93g, oleic acid 22.60g.

4.0 g of commercially available lipase preparation (derived from Candida) was added to p
The mixture was placed in a 1000ml phosphate buffer solution of 1+7.3 liters and subjected to the same procedure as in Example 1 to obtain 18.34 g of sorbitan oleate.

Patent Applicant: Kogyo Seiyaku Co., Ltd., Hajime Seino, Takeshi Uchihori, Patent Attorney, Patent Attorney Takashi Akaoka “Dog゛□・ ゝ・・・ Contents of Amendment Procedural Amendment Written October 26, 1980 Commissioner of the Japan Patent Office 2 , Name of the invention Process for producing sugar alcohol fatty acid ester 3, Relationship with the case of the person making the amendment Patent applicant name (350) Daiichi Kogyo Seiyaku Co., Ltd. (and 2 others)
4. Agent 5. Date of amendment order Voluntary 6. Number of inventions increased by amendment None 7. Claims subject to amendment, Detailed explanation of the invention 8. Contents of amendment TT Go; Lee Mi, amend the claims as follows.

[A method for producing a sugar alcohol fatty acid ester using an enzyme, which comprises incubating sorbitol or sorbitan and a higher fatty acid in the presence of a hydrolase.

2. Insert the following between page 3, line 15 and line 16 of the specification.

[Among them, Candida Cylindrace
Lipase MY (manufactured by Meito Sangyo @) derived from A has a significantly high production yield of sugar alcohol ester. ” 1st 11th 1st FL ↑ll'l 11-1st [February 1, 1985 1, Display of the case 1984 Patent Application No. 084777 2, Name of the invention Process for producing sugar alcohol fatty acid ester 3. Relationship with the case of the person making the amendment Patent applicant name (350) Dai-ichi Kogyo Seiyaku Co., Ltd. (two idiots)
4. Agent's own Invention Specification Document 1. Name of the invention Process for producing sugar alcohol fatty acid ester 2. Claims Characterized by incubating sorbitol or sorbitan and a higher fatty acid in the presence of a hydrolytic enzyme. A method for producing sugar alcohol fatty acid ester using an enzyme.

3. Detailed Description of the Invention The present invention relates to a method for producing sugar alcohol fatty acid esters 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 dispersed as a microemulsion in the presence of a fatty acid alkali metal salt. A microemulsion method in which a sugar and a fatty acid ester are reacted by melting them together with a fatty acid alkali metal salt, and a direct method 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 first discovered a method for producing sugar fatty acid esters from sugars and higher fatty acids using hydrolytic enzymes such as lipase, as a method of conducting the reaction at low temperatures and in an aqueous system. Ta. We have discovered that this can be further applied to sugar alcohols, leading to the present invention.

The present invention resides in a method for producing a sugar alcohol fatty acid ester using an enzyme, which comprises incubating sorbitol or sorbitan and a higher fatty acid in the presence of a hydrolase.

Sugar alcohol components that can be used in the present invention include sorbitol and sorbitan.

Suitable higher fatty acids are saturated or unsaturated fatty acids having 8 to 22 carbon atoms. Examples include:

caprylic acid, capric acid, lauric acid, myristic acid,
Palmitic acid, stearic acid, arachidic acid, behenic acid, caproleic acid, Linderic acid, myristoleic acid, palmitoleic acid, oleic acid, cadreic acid, erucic acid, decadienoic acid, linoleic acid, hiragic acid, lylunic acid, eicosatrienoic acid , docosatrienoic acid, hexadecatetraenoic acid, stearidonic acid, arachidonic acid, docosatetraenoic acid, eicosapencitric acid, sardine acid, sabinic acid, ibrolic acid, yarabinolic acid, ricinoleic acid, feronic acid.

As the enzyme, a hydrolytic enzyme, especially a lipase, is used. As is well known, there are lipases of animal origin and those of microbial origin; either of these may be used. For example, those derived from pig pancreas and those derived from microorganisms,
gill++s, Rh1zopus, Pseu-d
omonas, Enterobacterium, Ch.
romobacterium.

Geotrichum, Penicillium,
Some are derived from microorganisms such as Mucor and Candida. These enzymes do not necessarily need to be isolated and used; for example, as a crude enzyme such as pancreatin,
Alternatively, commercially available enzyme preparations containing lipase can be used as is.

Among them, Candi, da Cylindrace
1ipase MY (manufactured by Meito Sangyo @) derived from A has a significantly high production yield of sugar alcohol ester.

The optimum pH for these enzymes is between 5 and 8, but p114
to 9 pi (range may be used.

The reaction is carried out by adding the substrate and enzyme to a buffer and incubating at 20 to 60°C, preferably 30 to 50°C, until equilibrium is reached.

The ratio of sugar alcohol and fatty acid is selected in the range of 6:1 to 1:6 (weight ratio), the total substrate concentration is 1 to 30%,
Generally, a few percent is used. 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.

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 can be stopped in this state, and the 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 Commercially available lipase preparation (derived from Candida)2. OOg.

Add 3.64 g of sorbitol and 22.56 g of oleic acid to 1000 mJ2 of phosphate buffer at pH 5.4,
72 at 40℃ while stirring in a magnetic cooler.
Incubated for hours.

The reaction mixture is freeze-dried, the resulting freeze-dried product is extracted with chloroform, and the extract is concentrated under reduced pressure.

Dissolve the chloroform extract in tetrahydrofuran and add 30
Centrifuge at 00 r, p, m and separate into tetrahydrofuran soluble and tetrahydrofuran insoluble fractions.

Gel permeation chromatography was performed on the tetrahydrofuran soluble fraction, and the fraction eluting as the first peak was collected, and sorbitol oleate 7,
49 g was obtained.

Example 2 The same procedure as in Example 1 was repeated except that 22.8 g of stearic acid was used instead of 22.56 g of oleic acid to obtain 6.27 g of sorbitol stearate.

Example 3 Sorbitol 5.46g, oleic acid 22.56g.

4.0 g of commercially available lipase preparation (derived from Candida) was added to p
The mixture was placed in a 1000ml H7,3 phosphate buffer solution and incubated at 40°C for 72 hours while stirring with a Magsothysox stirrer. Thereafter, the same treatment as in Example 1 was carried out to obtain 16.92 g of sorbitol oleate.

Example 4 Sorbitan 3.28g, oleic acid 22.56g.

Commercially available lipase preparation ((derived from: Hdida) 2.0g
The mixture was placed in a phosphate buffer solution of H7,3 i o o o, t7, and the same procedure as in Example 1 was performed to obtain 8.65 g of sorbitan oleate.

Example 5 Sorbitan 4.93g, oleic acid 22.60g.

4.0 g of commercially available lipase preparation (derived from Candida) was added to p
i (7,3) in 1000 d of phosphate buffer solution, and the same procedure as in Example 1 was carried out to obtain 18.34 g of sorbitan oleate.

Patent applicant Dai-Kogyo Seiyaku Co., Ltd. Hajime Kiyono Tsuyoshi Hori Agent: Patent attorney Taku Akaoka-husband-1) '7,17 pa 2'-no

Claims (1)

[Claims] A method for producing a sugar alcohol fatty acid ester using an enzyme, which comprises kincubating sorbitol or sorbitan and a higher fatty acid in the presence of a hydrolase.