JP2000212588A - Production of highly unsaturated fatty acid-containing glyceride - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a method for producing a highly unsaturated fatty acid- containing glyceride by which the glyceride containing both eicosapentaenoic acid and docosahexaenoic acid concentrated so as to be highly pure is produced at a low cost with high efficiency. SOLUTION: This method for producing a glyceride containing >=30% eicosapentaenoic acid and >=20% docosahexaenoic acid comprises using an oil and fat containing >=20% eicosapentaenoic acid and >=10% docosahexaenoic acid, obtained by mixing an oil and fat derived from a shellfish containing >=30% eicosapentaenoic acid, and an oil and fat derived from a fish containing >=20% docosahexaenoic acid in an arbitrary rate.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a method for producing glycerides containing polyunsaturated fatty acids, and more particularly to a method for producing glycerides containing polyunsaturated fatty acids in which both eicosapentaenoic acid and docosahexaenoic acid are concentrated with high purity.

[0002]

2. Description of the Related Art Lipids of fish (fish oil) such as sardines, mackerel, saury and horse mackerel, or lipids of algae such as red algae and brown algae,
Fatty acids such as lipids of crustaceans, shellfish, and marine animals contain a large amount of highly unsaturated fatty acids (hereinafter, referred to as PUFAs).

Among them, eicosapentaenoic acid (hereinafter, referred to as eicosapentaenoic acid)
EPA) and docosahexaenoic acid (hereinafter, referred to as DHA) are unsaturated fatty acids of the ω-3 series, and their physiological activities have attracted attention in relation to prostaglandins and thromboxanes.

[0004] EPA is already marketed as a drug for arteriosclerosis and hyperlipidemia, and DHA has a characteristic physiological activity different from that of EPA such as an action of improving the retinal reflex function and an action of improving the memory and learning function. Is recognized,
Development in the pharmaceutical and food fields is underway.

[0005] Methods for concentrating PUFA mainly composed of EPA and DHA as described above include (1) chromatography, (2) urea addition, (3) low-temperature solvent fractional crystallization, and (4) molecular distillation. Or a vacuum distillation method, (5) a liquid-liquid partition method, (6) a method using an adduct to a double bond, (7)
A supercritical extraction method, a method using (8) lipase, a method combining these methods, and the like are known.

[0006] In general, PUFAs in pharmaceuticals are mainly used in the form of fatty acid ethyl esters concentrated to high purity. Fatty acid ethyl esters are glycerides (monoacylglycerol, diacylglycerol, triacylglycerol) due to their absorption in the body. Is defined to be inferior). On the other hand, PUFAs used in the food field, even in the form of glyceride, are generally products in which only one of EPA and DHA is concentrated, or products containing both in low purity. .

Therefore, glycerides having both EPA and DHA physiological activities, that is, EPA and DHA, will be
Glycerides containing both high purity are expected and establishment of an inexpensive production method thereof is being demanded.

[0008]

However, when tuna oil or sardine oil or the like, which is generally used as a raw material, is used alone by an existing technique such as a molecular distillation method or a hydrolysis method using lipase, EPA and DHA are used. However, it is very difficult to efficiently obtain highly pure glyceride, and even if it is obtained, it is very expensive in terms of yield and the like.

Accordingly, an object of the present invention is to provide a method for producing a highly unsaturated fatty acid-containing glyceride capable of efficiently and inexpensively producing a glyceride in which both EPA and DHA are concentrated with high purity. I do.

[0010]

Means for Solving the Problems To achieve the above object, the present invention relates to a highly unsaturated fatty acid-containing glyceride for producing a glyceride in which highly unsaturated fatty acids are concentrated using a highly unsaturated fatty acid-containing fat as a raw material fat. In the production method, the raw material fat is mixed at an arbitrary ratio between a shellfish-derived fat containing 30% or more of eicosapentaenoic acid (area% in constituent fatty acids, the same applies hereinafter) and a fish-derived fat containing 20% or more of docosahexaenoic acid. The mixed fats and oils contain eicosapentaenoic acid and docosahexaenoic acid in an amount of 20% or more and 10% or more, respectively.

The main raw materials for providing PUFAs include fish, marine animals such as whales, shellfish, algae, microorganisms, etc., and all of them contain only one of EPA and DHA. Alternatively, both contain only low purity and are not suitable as raw materials for producing glycerides in which both are concentrated.

However, according to the present invention, EPA and DH can be prepared by mixing the above fats and oils in advance at an arbitrary ratio.
A containing at least 20% and at least 10% of A can be obtained. If the said fat is used as a raw material, EPA and DH can be obtained.
It is possible to efficiently and inexpensively produce glyceride in which A is highly concentrated.

That is, the present invention relates to an oil or fat containing EPA with high purity, preferably a scallop oil containing EPA at least 30% and an oil or fat containing DHA with high purity, preferably DHA.
Of tuna or bonito oil containing at least 20% by weight of EPA and DHA, respectively.
EPA using oils and fats containing at least 10% and at least 10%
And DHA both efficiently and inexpensively produce glyceride concentrated with high purity.

As a method for concentrating glyceride in the present invention, any existing method such as hydrolysis with lipase, solvent fractionation, molecular distillation and the like may be used.

The lipase used for the hydrolysis in the present invention includes genus Candida, genus Alcaligenes, genus Mucor, genus Rhizopus,
Examples include lipases derived from microorganisms such as Pseudomonas and Geotricum, and lipase derived from pig pancreas.
More preferably PUF including EPA and DHA
Hardly acts on the binding between A and glycerin, ie, PU
Lipases with as low a substrate specificity as possible for FA are preferred, and in particular, Candida cylindracee (Candi).
A lipase derived from D. cylindracea is preferred.

The conditions for the hydrolysis reaction of lipase used in the present invention are the same as those for ordinary lipase. That is, the amount of lipase used is in the range of 10 to 2000 units (U), preferably 50 to 600 units (U), per raw material fat.

The amount of water to be added is in the range of 5 to 500% by weight, preferably 10 to 20% by weight, based on 1 g of raw material fat.
0% by weight.

The pH is preferably in the range of 3.0 to 8.5, and it is more effective to use a buffer to adjust the pH, and the pH is particularly preferably in the range of 4.0 to 7.5.

In order to perform an even more effective reaction,
An emulsifier, for example, polyvinyl alcohol, can also be used, and the addition of a bile salt is also effective for increasing the hydrolysis activity.

The reaction may be carried out in the atmosphere,
When a large amount of is contained, the deterioration of the fatty acid can be prevented by using an inert gas such as nitrogen gas. Further, an antioxidant such as tocopherol may be used in combination.

The hydrolysis reaction is preferably in the range of 20 to 60 ° C. If the temperature is lower than 20 ° C., the reaction is slow, and if it is higher than 60 ° C., nitrogen is deactivated. It is desirable to carry out in the range of 25 to 50 ° C.
The reaction is desirably agitated, but the reaction can be carried out in an emulsified state. Further, the reaction may be a batch type reaction, but an immobilized enzyme column can be used as a continuous type.

The degree of hydrolysis can be determined by sampling the hydrolyzed oil during the reaction and measuring the acid value. The content and yield of EPA and DHA in the obtained glyceride can be estimated by the degree of decomposition of the hydrolyzed oil, that is, the acid value of the hydrolyzed oil.

For the purpose of the present invention, it is desirable to terminate the reaction when the acid value of the hydrolyzed oil reaches 50 to 150. Usually, the reaction is carried out for a period of 1 to 24 hours.

If the acid value does not reach the target value,
It can be adjusted by reaction time and reaction temperature.

The hydrolyzed oil contains E, which is the target substance,
In addition to the glycerides containing a high content of PA and DHA, they contain free fatty acids generated during the reaction, and therefore need to be removed. In order to remove free fatty acids, in addition to the alkali deoxidation method and the molecular distillation method which are usually performed, a solvent extraction method, an ion exchange resin method, a low-temperature crystallization method and a reduced pressure steam distillation method, or a method combining these methods is used. Can be applied.

In addition to the above-described lipase concentration method, there is a solvent fractionation method (see Japanese Patent Application Laid-Open No. 58-15598).

As the solvent used in the present invention, an organic solvent such as acetone or hexane may be used alone, or a mixture thereof in an optional ratio may be used. Also, lower alcohols such as methanol and ethanol, water, etc.
You may mix about 0-20 weight% with respect to the above-mentioned solvent. The solvent can be recovered and reused later. The amount of solvent depends on the type of solvent, crystallization temperature, amount of crystallization, and required P
Although it depends on the UFA concentration, it is preferably used in an amount of 1 to 20 times the amount of fat or oil. The crystallization temperature is at least 5 ° C or lower, preferably -5 to -40 ° C.

The above series of operations may be performed in one step, or may be performed stepwise with a temperature gradient.

PUFA can also be concentrated by the above-mentioned molecular distillation method (see JP-A-9-57685).

For example, a falling-film molecular distillation apparatus or a centrifugal molecular distillation apparatus can be used.

At this time, the degree of vacuum is preferably 0.1 mmHg or less, and the temperature of the evaporation surface is 150 to 35.
The heat treatment may be performed at 0 ° C., preferably at 230 to 300 ° C.

Here, the higher the degree of vacuum, the lower the temperature of the evaporating surface can be processed. Therefore, in order to avoid the risk of thermal denaturation of the product, it is desirable to increase the degree of vacuum as much as possible and to process at a low temperature. .

In any case, in the molecular distillation method or the reduced pressure distillation method, in addition to the degree of vacuum and the evaporation surface temperature, the processing speed, the type of coexisting substance, the structure of the apparatus, etc. are related.
Since it is influenced by the quality, the type, structure and distillation conditions of the apparatus are not limited by the above contents.

[0034]

DESCRIPTION OF THE PREFERRED EMBODIMENTS Hereinafter, embodiments of the present invention will be described, but the present invention is not limited to these embodiments.

[0035]

Example 1 Scallop oil (EPA: 36.0%, DHA:
8.2%, acid value: 4.6) and bonito oil (EPA: 6.1)
%, DHA: 27.5, acid value: 0.5) were previously mixed at a ratio of 3: 1 (EPA: 27.2).
%, DHA: 13.3%, acid value: 3.0)
100 g of water containing 1.0 g of lipase powder (6,000 units / g) derived from the genus Candida
Was added, and an enzymatic reaction was carried out with stirring at 40 ° C. and pH 7.0 for 24 hours to obtain a hydrolyzed oil.

Next, the hydrolyzed oil was centrifuged, and after collecting the oil layer, glycerin was removed by washing with water.
Subsequently, using a falling film molecular distillation apparatus, the degree of vacuum was set at 0.
005mmHg, evaporation surface temperature 200 ° C, flow rate 30g / L
To remove free fatty acids,
And 50.5 g of glyceride in which DHA was concentrated.

Glyceride has an acid value of 1.1 and EPA
And DHA were 33.1% and 22.0%, respectively. The fatty acid composition is as shown in Table 1.

[0038]

[Table 1]

[0039]

Example 2 Scallop oil (EPA: 36.0%, DHA:
8.2%, acid value: 4.6) and tuna oil (EPA: 7.1)
%, DHA: 27.6%, acid value: 0.2) were previously mixed at a ratio of 1: 2, and the obtained fat (EPA: 17.2%) was obtained.
2%, DHA: 20.5%, acid value: 1.8) 100 g
1.0 g of lipase powder (30,000 units / g) derived from the genus Alcaligenes
Was added thereto, and an enzymatic reaction was carried out with stirring at 40 ° C. and a pH of 7.0 for 24 hours to obtain a hydrolyzed oil.

The oil layer was recovered and glycerin and free fatty acids were removed in the same manner as in Example 1 to obtain 39.5 g of glyceride in which EPA and DHA were concentrated. The acid value of glyceride is 1.0, and EPA and DHA are each 35.4.
% And 21.3%. The fatty acid composition is as shown in Table 2.

[0041]

[Table 2]

[0042]

Example 3 Scallop oil (EPA: 36.0%, DHA:
8.2%, acid value: 4.6) and tuna oil (EPA: 7.1)
%, DHA: 27.6%, acid value: 0.2) were previously mixed at a 2: 1 ratio (EPA: 27.1%).
%, DHA: 14.9%, acid value: 3.0) 100 g,
Using a falling film molecular distillation apparatus, vacuum degree is 0.005m
Treatment was performed under the conditions of mHg, evaporating surface temperature of 290 ° C., and flow rate of 50 g / L, to obtain 31.2 g of glyceride enriched in EPA and DHA as a residue. The acid value of glyceride is 1.0
EPA and DHA are 30.3% and 2 respectively.
0.1%. The fatty acid composition is as shown in Table 3.

[0043]

[Table 3]

[0044]

Comparative Example 1 Scallop oil (EPA: 36.0%, DHA:
8.2%, acid value: 4.6) and 100 g of lipase powder (6,000) derived from the genus Candida.
(Unit / g) 100 g of water containing 1.0 g is added, and 40
An enzymatic reaction was carried out while stirring at 24 ° C. and pH 7.0 for 24 hours to obtain a hydrolyzed oil.

The oil layer was recovered and glycerin and free fatty acids were removed in the same manner as in Example 1 to obtain 25.6 g of glyceride in which EPA and DHA were concentrated. The acid value of glyceride is 0.9, EPA and DHA are 38.8 each.
% And 16.6%. The fatty acid composition is as shown in Table 1.

[0046]

Comparative Example 2 Tuna oil (EPA: 7.1%, DHA: 2)
7.6%, acid value: 0.2) 100 g of lipase powder (3) derived from the genus Alcaligenes.
100 g of water containing 1.0 g of (0.00 units / g) was added, and an enzymatic reaction was carried out under stirring at 40 ° C. and pH 7.0 for 24 hours to obtain a hydrolyzed oil.

The oil layer was recovered and glycerin and free fatty acids were removed in the same manner as in Example 1 to obtain 24.6 g of glyceride in which EPA and DHA were concentrated. The acid value of glyceride is 1.1, EPA and DHA are each 14.8.
% And 26.9%. The fatty acid composition is as shown in Table 2.

[0048]

As described above, the present invention relates to a method for producing a highly unsaturated fatty acid-containing glyceride in which a highly unsaturated fatty acid-enriched glyceride is produced by using a highly unsaturated fatty acid-containing fat or oil as a raw fat or oil. Fats and oils,
A shellfish-derived fat containing 30% or more of eicosapentaenoic acid and a fish-derived fat containing 20% or more of docosahexaenoic acid are mixed at an arbitrary ratio, and eicosapentaenoic acid and docosahexaenoic acid are mixed in the mixed fat and oil in 20% or more and 10%, respectively.
% Or more, so that EPA and DHA can be efficiently and inexpensively produced with high-purity concentrated glycerides.

 ──────────────────────────────────────────────────続 き Continuing on the front page (72) Inventor Koji Nishioka 95-7 Minoki-cho, Fukuyama City, Hiroshima Prefecture Inside Ikeda Shokuken Co., Ltd. F-term in SHIKEN CO., LTD. (Reference) 4H059 BA33 BB05 BB07 BC06 BC48 CA02 CA13 CA20 CA38 CA51 CA99 EA17 EA21

Claims (4)

[Claims] 1. A method for producing a highly unsaturated fatty acid-containing glyceride, wherein a highly unsaturated fatty acid-enriched glyceride is produced by using a highly unsaturated fatty acid-containing fat as a raw fat, wherein the raw fat contains eicosapentaenoic acid in an amount of 30% or more. A mixture of shellfish-derived fats and fish-derived fats containing 20% or more of docosahexaenoic acid in any proportion, and containing 20% or more and 10% or more of eicosapentaenoic acid and docosahexaenoic acid in the mixed fats and oils, respectively. A method for producing a highly unsaturated fatty acid-containing glyceride, comprising: 2. The highly unsaturated fatty acid-containing glyceride according to claim 1, wherein the glyceride enriched in the polyunsaturated fatty acid contains 30% or more of eicosapentaenoic acid and 20% or more of docosahexaenoic acid. Manufacturing method. 3. The method for producing a highly unsaturated fatty acid-containing glyceride according to claim 1, wherein the shellfish-derived fat or oil is a scallop-derived fat or oil. 4. The method for producing a highly unsaturated fatty acid-containing glyceride according to claim 1, wherein the fish-derived fat or oil is a skipjack or tuna-derived fat or oil.