JPS6312289A - Production of gamma-linolenic acid glyceride - Google Patents

Abstract

PURPOSE:To obtain the titled substance utilizable as pharmaceuticals, etc., in high efficiency, by partially hydrolyzing an oil or fat containing GLA as a constituent fatty acid with an oil or fat hydrolase and removing free fatty acid from the product. CONSTITUTION:An oil or fat containing GLA (gamma-linolenic acid), e.g. seed oil such as RURI-JISHA seed oil, oil of algae belonging to Spirulina genus, etc., or oil of microorganism belonging to Cunninghamella genus is hydrolyzed by using preferably 50-500IU of a lipase originated from microorganism, animal or vegetable based on 1g of the oil or fat. The decomposition is usually carried out by mixing a GLA-containing oil or fat with an aqueous solution of lipase and hydrolyzing in emulsified state. The amount of water is preferably 1-50pts. wt. per 1pt. of the oil or fat and the degree of hydrolysis is preferably 40-85%. The titled substance can be separated from free fatty acid e.g. by removing the acid with an alkali.

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial Field of Application] The present invention relates to a novel method for producing γ-linolenic acid glyceride.

[Conventional technology]

γ-Linolenic acid is 6,9,12-octadecatrienoic acid, and its physiologically active effects have been studied in relation to prostaglandins, thromboxane, etc., and it is attracting attention as a therapeutic and preventive drug for various adult diseases. has been done.

A method of fractionating glyceride is known as a method for producing unsaturated fatty acid glyceride, but in order to produce γ-linolenic acid glyceride from fats and oils containing γ-linolenic acid as a constituent fatty acid (hereinafter abbreviated as GLA-containing fats and oils), I can't get enough results.

In addition, after converting the raw material glyceride into fatty acids or lower alcohol esters of fatty acids, (a) chromatographic separation, (b) urea addition fractionation, (c) fractional distillation, and (c) liquid-liquid distribution. It is also possible to concentrate the glyceride using conventionally known techniques, such as the method described above, and then esterify it with glycerin to produce glyceride again, but this is not only a complicated process, but also produces isomers of unsaturated fatty acids. Not practical.

As a method of concentrating specific glycerides using fat hydrolase, long-chain polyunsaturated fatty acids (2
Attempts have been made to make glycerides of fatty acids (fatty acids having 0 or more carbon atoms and 3 or more double bonds) (Japanese Unexamined Patent Application Publication No. 1983-1999)
-165796, JP-A-61-15692 and 1
No. 5693).

However, there has been no report on glyceride of γ-linolenic acid having 18 carbon atoms.

As described above, no effective method is known for producing γ-lyl/acid glyceride in the form of glyceride.

[Problem that the invention seeks to solve]

An object of the present invention is to provide a method for efficiently and effectively producing γ-linolenic acid glyceride from GLA-containing oil or fat in the form of glyceride.

[Failure to solve the problem]

The present inventors investigated the difference in the degree of action of fat and oil hydrolase (hereinafter referred to as lipase) depending on the position of the unsaturated double bond in fatty acids, and found that lipase can , saturated fatty acids, oleic acid (9-octadecenoic acid), linoleic acid (9,12- octadecadienoic acid), α-linolenic acid (9,12,15
-octadecatrienoic acid) etc. and glycerin, but has little effect on the ester bond between γ-linolenic acid, which has an unsaturated bond at the Δ6 position, and glycerin. They discovered this and completed the present invention.

That is, the present invention is a method for producing γ-linolenic acid glyceride, which is characterized by partially hydrolyzing fats and oils containing γ-linolenic acid as a constituent fatty acid using an oil-fat hydrolase, and then removing free fatty acids.

In the present invention, GLA-containing oils and fats include seed oils such as evening primrose seed oil, saxifrage seed oil, and borage seed oil, oils and fats obtained from algae such as Spirulina, and oils and fats obtained from algae such as Caningamela, Phycomyces, Silpeltera, and Tamnibuium. Examples include oils and fats obtained from microorganisms.

In the present invention, the partial hydrolysis of GLA-containing fats and oils
A method of mixing A-containing fat and oil with a lipase aqueous solution and hydrolyzing it in an emulsified state, a method of floating oil droplets of GLA-containing fat in a lipase aqueous solution and hydrolyzing the mixture, and a method of hydrolyzing GLA-containing fat and oil by floating a porous membrane between the GLA-containing fat and lipase aqueous solution. This can be carried out by any method capable of hydrolyzing GLA-containing fats and oils with lipase, such as by contacting the GLA-containing fat with a lipase.

Lipases used in the present invention can be derived from microorganisms, animals, and plants. A specific example is Rhizopus chinensis.
lipase from Rhizopus chinensts
Delemer (Rhiz.

lipase derived from Candtda cyjin
dra cea), a lipase derived from Pseudomonas
Fluorescence (p36udom.

lipase derived from nas futuorescens),
G e o t ri
lipase from Penicillium candidum;
cyctopium)-derived lipase, mucor
Lipase from Mucor m1ehe1, Aspergillus nikah
Examples of lipases derived from microorganisms include lipases derived from microorganisms such as lipases derived from A. niger; examples of lipases derived from animals include pancreatic lipases from humans, pigs, and other animals; and examples of lipases derived from plants include castor seed lipases. Can be mentioned.

The amount of lipase used in the present invention varies depending on the type of lipase used, the hydrolysis method, and the fat or oil to be hydrolyzed, but is usually 10 to 1 for 12 fats and oils.
000 International Units (IU), preferably 50-500 IU
It is. The international unit (IU) is defined as the activity of an enzyme with lipase activity that liberates 1 micromole of fatty acids per minute when it hydrolyzes olive oil under optimal reaction conditions (temperature, pH). It is.

The amount of water used to perform the partial hydrolysis of the present invention is
Although it varies depending on the hydrolysis method, in the method of mixing a normal GLA-containing fat and oil with a lipase aqueous solution and hydrolyzing it in an emulsified state, the amount is 0.5 to 100 M times, preferably 1 to 50 times the weight of the fat. It is.

In the production of γ-linolenic acid glyceride of the present invention, the concentration and yield of the target unsaturated fatty acid are influenced by the hydrolysis rate of the raw material fat. This hydrolysis rate varies greatly depending on the unsaturated fatty acid concentration in the raw material oil, but is usually 4.
A hydrolysis rate of 0 to 85% is preferred.

The separation of γ-linolenic acid glyceride obtained by the partial hydrolysis of the present invention from free fatty acids can be carried out by methods such as alkaline deacidification, molecular distillation, steam distillation, ion exchange resin, and solvent extraction. ,
Alternatively, it can be carried out in the usual manner by a combination of these methods.

〔Effect of the invention〕

By using the method of the present invention, γ-linolenic acid glyceride can be effectively produced with high yield from GLA-containing fats and oils.

Further, the γ-linolenic acid glyceride obtained by the present invention can be used in pharmaceuticals, cosmetics, foods, etc. due to its physiologically active action.

〔Example〕

The present invention will be explained by examples.

Note that the hydrolysis rate was calculated using the formula shown below.

AVo: Acid value of raw material oil AV+: Acid value after hydrolysis Sv: Saponification value of raw material oil Example 1 Borage seed oil (acid value 0.8, saponification value 197°6,
γ-linolenic acid content 20.2U) 100S', lipase derived from Candida cylindracea (360U
/ Cape) 551N! Ion-exchange water 1002 contained in the mixture was added, and hydrolysis was carried out with stirring at 30° C. for 18 hours. After the reaction was completed, the oil layer was collected to obtain a hydrolyzed oil. The acid value of this hydrolyzed oil is 143°2, and the hydrolysis rate is 711.
%Met.

This hydrolyzed oil was neutralized with an aqueous solution of sodium hydroxide, and the soapy water was removed using a centrifuge to obtain glycerides of 2&11. The yield of γ-linolenic acid glyceride is 2 & 1
The acid value was 0.2.

In addition, γ-linolenic acid in glyceride is
．． It was 3 times more concentrated. The fatty acid composition of this glyceride is shown in Table 1.

Example 2 Borage seed oil was hydrolyzed for 41 hours in the same manner as in Example 1. The acid value of the obtained hydrolyzed oil was 160.0, and the hydrolysis rate was 80.6%.

This hydrolyzed oil was treated in the same manner as in Example 1 to obtain 19.2
9 glycerides were obtained. The yield of γ-linolenic acid glyceride was 19.2%, and the acid value was 0.1.

γ-linolenic acid in the glyceride was 21 times more concentrated than in the raw oil. The fatty acid composition of this glyceride is summarized in Table 1.

Example 3 Borage heel oil was hydrolyzed for 1 hour in the same manner as in Example 1. The acid value of the obtained hydrolyzed oil was 100.2, and the hydrolysis rate was 5083%.

This hydrolyzed oil was treated in the same manner as in Example 1, and 49,7
Glyceride of t was obtained. The yield of γ-linolenic acid glyceride was 49.7%, and the acid value was 0.1.

The r + lylunic acid in the glyceride was concentrated to ZO times that of the raw material oil. The fatty acid composition of this glyceride is shown in Table 1.

Example 4 Ion-exchanged water 3002 containing 7.7 F of lipase derived from Rhizopus delemer (6500 IU/V) was added to the same oil 100 F as in Example 1, and hydrolysis was performed while stirring at 30° C. for 24 hours. The acid value of the obtained hydrolyzed oil was 87.
．． 4, and the hydrolysis rate was 4&8%.

This hydrolyzed oil was treated in the same manner as in Example 1 to obtain 55,3
Glyceride of F was obtained. The yield of γ-linolenic acid glyceride was 55.3%, and the acid value was 0.2.

The γ-linolenic content in the glyceride was 1.8 times more concentrated than in the feedstock oil. The fatty acid composition of this glyceride is shown in Table 1.

Example 5 Lipids were extracted from commercially available Spirulina tablets 6009 using a mixed solvent of chloroform and methanol, and separated by silica gel column chromatography to obtain glyceride 265'.

This glyceride (acid value 0.1, saponification value 194.1, γ
- Linolenic modified 5.5%) 20 f, lipase derived from Candida cylindrace (360 IU/rr
q) 50 f of ion-exchanged water containing ssη was added, and hydrolysis was performed while stirring at 30°C for 8 hours.

After the reaction was completed, the oil layer was collected to obtain a hydrolyzed oil.

The acid value of this hydrolyzed oil was 171.8, and the hydrolysis rate was 88.5.

This hydrolyzed oil was treated in the same manner as in Example 1, and
Glyceride of f was obtained. The yield of γ-linolenic acid glyceride was 11.2%, and the acid value was 0.1.

γ-linolenic acid was 1.7 times more concentrated than the raw material oil. The fatty acid composition of this glyceride is shown in Table 1.

Comparative Example 1 Flaxseed oil (acid value 1.2, saponification value 195.7, α-linolenic acid 56.2%) 1009 was treated under the same conditions as Example 1.
．． Hydrolysis was carried out for 5 hours. The acid value of the obtained hydrolyzed oil was 138.6, and the hydrolysis rate was 70.2.

This hydrolyzed oil was treated in the same manner as in Example 1, resulting in 30.4
5' glyceride was obtained. The yield of glyceride is 30.4
%, and the acid value was 0.1. The fatty acid composition of this glyceride was as shown in Table 1, and α-linolenic acid was not concentrated.

Comparative example 2 Hydrolysis was carried out in the same manner as in Comparative Example 1 for 2 hours.

The acid value of the obtained hydrolyzed oil was 157.1, and the hydrolysis rate was 79.7%.

This hydrolyzed oil was treated in the same manner as in Example 1 to obtain 21, 4
Glyceride of F was obtained. The yield of glyceride is 21.4
%, and the acid value was 0.1. The fatty acid composition of this glyceride was as shown in Table 1, and Q + IJ phosphoric acid was not concentrated.

Examples 6 to 9 9) Hydrolysis on the same side as in Example 1 was carried out using 20 g of the same oil as in Example 1.

The obtained hydrolyzed oil was treated in the same manner as in Example 1 to recover glyceride.

The hydrolysis rate, glyceride and γ-linolenic acid recovery rates are summarized in Table 2.

Example 10 To the same oil LOOP as in Example 1, 1002 ion-exchanged water containing 688 q of lipase derived from Candida cylindracea (360 IU/■) was added, and hydrolysis was carried out with stirring at 30° C. for 24 hours.

The oil layer was collected to obtain hydrolyzed oil. The acid value of this hydrolyzed oil was 183.9, and the hydrolysis rate was 93.1.

This hydrolyzed oil was treated in the same manner as in Example 1 to obtain 8.02%
of glycerides were obtained. The yield of γ-linolenic acid glyceride is 8. Q%, and the acid value was 0.2.

1-linolenic acid in guxeride was concentrated 1.9 times as much as the raw material oil. The results are summarized in Table 2.

As is clear from the above results, by using the method of the present invention, γ-linolenic acid glyceride can be efficiently produced.

Claims (1)

[Claims] 1. A method for producing γ-linolenic acid glyceride, which comprises partially hydrolyzing an oil or fat containing γ-linolenic acid as a constituent fatty acid using an oil-fat hydrolase, and then removing free fatty acids.