KR20060106161A - A supplementary food composition comprising reformed triglyceride lipid - Google Patents

Abstract

The present invention relates to an arachidonic acid-source and a food supplement composition for inhibiting in vivo side effects of arachidonic acid, characterized by comprising a reconstituted triglyceride represented by the following Chemical Formula 1:

Formula 1

In the above formula, at least one of R ₁ , R ₂ and R ₃ is arachidonic acid and at least one is gamma-linolenic acid.

The food supplement composition of the present invention helps the positive metabolism of arachidonic acid in the application of arachidonic acid to baby food, and at the same time minimizes the possibility of side effects of inflammatory diseases due to the excess metabolism of arachidonic acid, and minimizes other lipids such as saturated fatty acids. One functional fatty acid composition can be supplied.

Reconstituted lipids, arachidonic acid, gamma-linolenic acid, triglycerides, food

Description

A Supplementary Food Composition Comprising Reformed Triglyceride Lipid

Figure 1 shows the human metabolism mechanism of the constituent material of the present invention arachidonic acid and gamma-linolenic acid.

The present invention relates to a food supplement composition comprising reconstituted triglyceride lipids, and more particularly, to a food supplement composition comprising lipids reconstituted with arachidonic acid and gamma-linolenic acid into a single form of triglyceride.

Arachidonic acid is distributed in the human body in a small amount compared to other fatty acids, but is an unsaturated fatty acid which is very essential for the growth and development of the infant and brain and nerve development.

Conventionally, arachidonic acid oil (Patent No. 3002036) or an unmodified form derived from microorganisms such as Mortierella alpina and phidium incidio islands or patented egg yolks and fish oils to sufficiently supply such arachidonic acid to infants. Arachidonic acid oil compositions have been used in combination as a supplement for baby food.

However, when arachidonic acid, which is a very essential nutrient for growing infants, is metabolized excessively, there is a fear that overexpression of inflammatory expression factor in infants' bodies. In other words, if arachidonic acid metabolites such as 4-series leukotrienes, platelet-activating factor (PAF), and 2-series prostaglandins are excessively produced and accumulated in the body, These metabolites may act as a major inflammatory factor in the expression of atopic dermatitis or rheumatoid arthritis (Johnson, et al., J. Nutr. 127: 1435-1444 (1997)), especially in adolescents of growing age. When the acid is excessively metabolized and accumulated, there is a high possibility of expressing or intensifying inflammatory immune diseases such as atopic dermatitis and rheumatoid arthritis, which are commonly found in infants. In particular, infants whose parents have a history of atopic dermatitis or rheumatoid arthritis, i.e., have a family history of inflammatory immune disease, are more inevitable.

Therefore, in order to adequately supply arachidonic acid, which is essential for human growth, and to suppress side effects in which arachidonic acid metabolites develop as inflammatory factors, omega-3 or omega that metabolizes competitively with arachidonic acid such as oleic acid, alphalinolenic acid, EPA, DHA, etc. -9 unsaturated fatty acids can be applied, but this method unilaterally inhibits the metabolism of arachidonic acid, which may interfere with the positive metabolism of arachidonic acid, which is essential for the growth and development of infants. In particular, omega-3 fatty acid-based alphalinolenic acid, EPA, etc. are nutrients that metabolize preferentially to enzymes called Δ6 and Δ5-desaturase than omega-6 unsaturated fatty acids such as arachidonic acid and gammalinolenic acid. Since it can unilaterally interfere with the metabolic action of arachidonic acid in competition with it, it may result in halving the positive metabolism of arachidonic acid, and thus it is not preferable to apply to a baby food ingested by arachidonic acid.

In order to avoid this problem, the precursor of arachidonic acid, which is essential for the growth and development of infants, is helpful for the positive metabolism of arachidonic acid, and at the same time, it can control the expression of inflammatory factors due to the overmetabolism of arachidonic acid. There are a method of applying linoleic acid, γ-linolenic acid, and dihomo-γ linolenic acid, which are the 6 series fatty acids. Among these, linoleic acid has a very low conversion rate to γ-linolenic acid and arachidonic acid in the Δ6-desaturated enzyme reaction, and its anti-inflammatory function is also ineffective, and consequently, it is difficult to help positive metabolism of arachidonic acid. -γ linolenic acid is a structurally very low stability unsaturated fatty acid which is not suitable for baby food applications, and there is little vegetable or animal fats containing dihomo-γ linolenic acid naturally there is a problem in the practical application.

On the other hand, γ-linolenic acid, which is a precursor of arachidonic acid and precursors such as DGLA, 15-HETrE and PGE-1, which are anti-inflammatory substances, is also known as evening primrose oil and borage seed oil. Since it is contained in vegetable seed oil such as), it is easy to produce commercially, and it is helpful for positive metabolism to arachidonic acid as a precursor of arachidonic acid, and also anti-inflammatory substances such as DGLA, 15-HETrE and PGE-1. Since it is metabolized as a precursor, it can be said that this γ-linolenic acid is more suitable for applying as a baby food supplement material that can suppress the inflammatory expression due to the overmetabolism of arachidonic acid. van Gool, et al. have argued that growing infants can benefit from atopic dermatitis by taking at least 103 mg of γ-linolenic acid per day ( Am. J. Clin. Nutr. 77: 943-951 (2003)), Fewtrell, et al. Have reported that γ-linolenic acid helps positive metabolism of arachidonic acid in growing infants ( J. Pediatr. 144: 471-9 (2004)).

The arachidonic acid-containing fats and oils used in conventional baby food supplements contain only about 2% of γ-linolenic acid, and only the unmodified arachidonic acid-containing fats are insufficient to obtain the above effects. As a method for applying such γ-linolenic acid, it was common to simply mix evening primrose seed oil or borage seed oil, but since the vegetable seed oil itself contains only a very small amount of γ-linolenic acid, a sufficient amount of γ-linolenic acid In order to apply to baby food, excess vegetable seed oil must be mixed, which causes infants to ingest excess glycerin, saturated fatty acids and other lipids in excess, and increase the amount of fatty acids consumed in a day, thus increasing infant nutritional imbalance. Problems may occur that may result. It is very undesirable to consume excessive amounts of fatty acids that are digested and absorbed in the small intestine by enzymes secreted by the pancreas. Therefore, reconstituted lipids in which a high content of vegetable seed oil origin γ-linolenic acid containing oils having a stronger potency of γ-linolenic acid in vegetable seed oil are combined with arachidonic acid and a triglyceride of a single form are more preferable in baby food.

On the other hand, the exclusion of arachidonic acid and its precursor γ-linolenic acid alone to be applied to infant food alone to expect a positive metabolism of arachidonic acid, gamma-linolenic acid is sufficient as arachidonic acid due to the limitation of △ 5-desaturating enzyme of the human body Since it is not converted, it is not preferable because it is less efficient than the direct application of arachidonic acid in the growth and development of the brain and brain and nerves.

The present inventors apply arachidonic acid directly to facilitate the positive metabolism of arachidonic acid in infants and at the same time to concentrate the precursor of gamma-linolenic acid in high content in order to suppress the side effects of inflammatory expression due to excessive metabolism of arachidonic acid. Therefore, the method of providing arachidonic acid and a single form of reconstituted lipid was considered as the most desirable. Finally, the present invention was completed by preparing a reconstituted triglyceride in which arachidonic acid and gamma-linolenic acid are present in a single form of triglyceride.

It is therefore an object of the present invention to provide a food supplement composition which is an arachidonic acid-source comprising reconstituted triglycerides and which can suppress the in vivo side effects of arachidonic acid.

Another object of the present invention is to provide a method for efficiently preparing reconstituted triglycerides.

Other objects and advantages of the present invention will become apparent from the following detailed description, claims and drawings.

According to one aspect of the invention, the present invention provides a food supplement composition for inhibiting in vivo side effects of arachidonic acid-source and arachidonic acid, characterized in that it comprises a reconstituted triglyceride represented by the following formula (1):

In the above formula, at least one of R ₁ , R ₂ and R ₃ is arachidonic acid and at least one is gamma-linolenic acid.

According to another aspect of the present invention, the present invention provides an efficient method for preparing the reconstituted triglyceride of the formula (1). The method of the present invention is largely carried out in two ways, one of which is a method using only (i) an esterification reaction, and the other is a method using simultaneously (ii) an esterification reaction and a transesterification reaction.

The method using only the esterification reaction in the above method includes the following steps: (a) hydrolyzing the microorganism-derived, egg yolk-derived or fish oil-derived arachidonic acid-linked triglyceride to obtain free arachidonic acid; And (b) adding lipase to the free arachidonic acid, gamma-linoleic acid, and glycerin mixture and reacting at a temperature of 50-65 ° C. and 10 ⁻¹ −10 ⁻³ torr to obtain a triglyceride combined with arachidonic acid and gamma-linoleic acid. Steps.

Among the above methods, the method of simultaneously using the esterification reaction and the transesterification reaction includes the following steps: (a) (i) microorganism-derived, egg yolk-derived or fish oil-derived arachidonic acid-linked triglycerides, (ii) Treating lipase with a mixture comprising glycerin and (iii) water to form mono-glycerides and diglycerides; And (b) adding gamma-linoleic acid to the reaction product of step (a) and reacting at a temperature of 50-65 ° C. and a pressure of 10 ⁻¹ −10 ⁻³ torr to obtain triglycerides in which arachidonic acid and gamma-linoleic acid are combined. step.

According to another aspect of the invention, the present invention is arachidonic acid-source characterized in that it comprises a reconstituted triglyceride prepared by the method of the present invention described above, food supplement composition for inhibiting in vivo side effects of arachidonic acid To provide.

The present inventors have applied arachidonic acid directly to facilitate the positive metabolism of arachidonic acid in infants and at the same time to suppress the side effects of inflammatory expression due to excessive metabolism of arachidonic acid to a high content of its precursor gamma-linolenic acid. It was conceived that a method of concentrating and providing arachidonic acid and a single form of reconstituted lipid was the most desirable. Finally, a reconstituted triglyceride was prepared in which arachidonic acid and gamma-linolenic acid were present in a single form of triglyceride.

Lipids reconstituted with arachidonic acid and gamma-linolenic acid in a single form of triglycerides according to the present invention provide arachidonic acid while suppressing the side effects of arachidonic acid (eg, causing atopic dermatitis). In particular, reconstituted triglycerides are applied to infant foods to help growth and brain / nerve development of growing infants, and to suppress side effects of inflammatory diseases such as atopic dermatitis and rheumatoid arthritis in infants due to excessive metabolism of arachidonic acid. In addition, the application of concentrated arachidonic acid and gamma-linolenic acid, which removes other lipids such as saturated fatty acids, can help regulate the absolute daily intake of fatty acids in infants, while at the same time taking sufficient amounts of arachidonic acid and gamma-linolenic acid. do.

In a preferred embodiment of the present invention, the content of the reconstituted triglyceride is 0.1-80% by weight, more preferably 10-80% by weight, even more preferably 50-75% by weight, based on the total composition, Most preferably 65-70% by weight.

According to a preferred embodiment of the present invention, in the reconstituted triglyceride arachidonic acid is at least 10%, gamma-linolenic acid is at least 5%, more preferably arachidonic acid is at least 20%, gamma-linolenic acid is at least 10% Even more preferably 20-30% of arachidonic acid, 10-20% of gamma-linolenic acid, most preferably 20-25% of arachidonic acid, and 10-15% of gamma-linolenic acid.

According to the present invention, the lipid of the arachidonic acid and gamma linolenic acid reconstituted into a single form of triglyceride can be prepared by selecting one of organic synthesis and enzyme synthesis. Organic synthesis has advantages in terms of productivity because of short reaction time and high reconstitution rate. However, the basic product or acid catalyst should be used depending on the type of lipid, and since most of the lipid is reconstituted at high temperature, the final product obtained through this process Is not preferred for application to food, especially baby food.

Thus, preferably, the reconstituted triglycerides of the present invention are prepared by enzymatic methods using lipases, wherein the microorganism-derived, egg yolk-derived or fish oil-derived arachidonic acid-linked triglycerides and vegetable seed oil-derived gamma-linoleic acid are used as lipases. It is prepared by reconstitution through the enzymatic method used. Details of the preparation method are described below.

According to the preparation method of the present invention, microorganism-derived, egg yolk-derived or fish oil-derived arachidonic acid-linked triglycerides are used as reactants, more preferably Mortierella alpina and Pythium. microorganism-derived arachidonic acid-linked triglycerides derived from insidiosum ), and most preferably microorganism-derived arachidonic acid-linked triglycerides derived from Mortierella alpina.

The gamma-linolenic acid used in the preparation method of the present invention is obtained by extracting and purifying from vegetable seed oil, preferably derived from vegetable seed oil derived from evening primrose seed oil, borage seed oil and blackcurrant oil, and most preferably Borage is derived from seed oil.

As the lipase used in the preparation method of the present invention, various lipases known in the art may be used, and preferably Candida rugosa , Candida antartica , Candida cylindracea ), Pseudomonas sp. , And Rhizomucor miehei -derived lipases, most preferably Candida Antartica and Rizomuco miehei -derived lipases. Such lipases can be obtained directly from the microorganisms described above, but the lipase gene is transformed into other microorganisms, such as Aspergillus oryzae , and obtained from cultures obtained by culturing the transformed microorganisms. It may be.

In the preparation method of the present invention, step (b) in which the esterification reaction, or the esterification reaction and the transesterification reaction occur simultaneously, is performed at 30-80 ° C., preferably at 50-65 ° C., and at a temperature of 10 ⁻¹ -10 ⁻ under reduced pressure. ^{Under 3} torr pressure, more preferably at 60 ° C. temperature and 10 ⁻³ torr pressure. The reaction time is generally 6-98 hours, preferably 10-70 hours, more preferably 20-60 hours, most preferably 40-55 hours. This reaction condition prevents the reaction of step (b) from proceeding to the hydrolysis reaction and proceeds to the esterification reaction and the transesterification reaction.

In the method of using the esterification reaction and the transesterification reaction in the production method of the present invention, step (a) is to form a mono-glyceride and di-glyceride in which one or two fatty acids are bonded to the glycerin skeleton Step. The mono-glycerides and di-glycerides are formed by hydrolysis of the arachidonic acid-linked triglycerides used as reactants, and may also be formed by binding fatty acids produced by hydrolysis to glycerine as a reactant. . That is, in step (a), a hydrolysis reaction and an esterification reaction occur simultaneously. For this reaction pattern to be achieved, water must be present as the reactant and the reaction conditions must be set appropriately. Preferred reaction conditions in step (a) are 30-40 ° C. temperature and atmospheric pressure (atmospheric pressure), different from step (b), more preferably 30-36 ° C. temperature and atmospheric pressure (atmospheric pressure).

The preparation method of the present invention using the esterification reaction and the transesterification reaction may be carried out by a process comprising the following steps: (a) (iii) gamma-linoleic acid-linked triglyceride, (ii) glycerin and ( Iii) treating the mixture comprising water to lipase to form mono-glycerides and diglycerides; And (b) adding arachidonic acid in the form of free arachidonic acid or ethyl ester to the reaction product of step (a) and reacting under a temperature of 50-65 ° C. and 10 ⁻¹ −10 ⁻³ torr to arachidonic acid and gamma-linoleic acid. Obtaining this bound triglyceride.

This method differs from the second method by (i) using free arachidonic acid or arachidonic acid in ethyl ester form instead of arachidonic acid in triglyceride form, and (ii) triglyceride in place of gamma-linoleic acid in free form. Form of gamma-linoleic acid. The remaining details of the method are the same as the second method.

Gamma-linoleic acid-linked triglycerides used in the present invention are vegetable seed oil, evening primrose seed oil, borage seed oil and blackcurrant oil, most preferably borage seed oil.

Free arachidonic acid or arachidonic acid in ethyl ester form for use in the present invention can be readily obtained from triglycerides through various methods known in the art. For example, free arachidonic acid can be readily obtained by hydrolysis (eg, hydrolysis by lipase) of arachidonic acid-linked triglycerides. Arachidonic acid in ethyl ester form, on the other hand, can be obtained by lipase treatment of arachidonic acid-linked triglycerides in the presence of ethanol, or by sulfuric acid treatment of free arachidonic acid.

According to the preparation method of the present invention, after the reconstituted triglyceride is prepared by the above steps, the lipase is removed by filtration, and the unreacted free fatty acid is removed by washing with a molecular distillation process, a silica column process or a weakly basic solution. If necessary, remove peroxides with activated charcoal or acid clay.

In the two production methods of the present invention, the second, that is, a method of simultaneously using the esterification reaction and the transesterification reaction in terms of yield and reaction progression and convenience, is preferred, which can be confirmed in the following examples. .

Lipids reconstituted with a single triglyceride of arachidonic acid and gamma-linolenic acid thus help the growth and brain / nerve development of growing infants, and atopic dermatitis and rheumatoid arthritis that may be caused by the excess metabolism of arachidonic acid. It can inhibit the onset or deepening of inflammatory diseases and provide their single form reconstituted lipids with higher purity of at least one of arachidonic acid or gamma linolenic acid to prevent excessive intake of glycerin, saturated fatty acids and other lipids. Applicable as an improved baby food supplement.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention in more detail, it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples in accordance with the gist of the present invention. .

Example 1

Containing 42% arachidonic acid derived from Mortierella alpina- 100 g of fatty acid fats hydrolyzed from fats and oils were prepared, and 100 g of 63% gamma linolenic acid in fatty acid form from Borage seed oil (Lipozen, Inc) Was prepared. The two components were mixed and stirred at 200 rpm at 60 ° C. with 30 g of glycerin in a vacuum flask equipped with a stirrer. After 10 minutes, 50 g of Candida antartica -derived lipase (Novozymes A / S) was added and reacted for 48 hours under reduced pressure of 10 ^-2 torr. When the product was analyzed by TLC, a single form of triglyceride reconstituted lipid was identified, and unreacted free fatty acid, mono- and diglycerides were removed through molecular distillation, and triglyceride combined with arachidonic acid and gamma linoleic acid. Reconstituted lipids were finally obtained. The composition of the mixture before the preparation of reconstituted lipids was analyzed by gas chromatography, and the results showed that arachidonic acid was 21.6% and gammalinolenic acid was 33.7%. After preparation of reconstituted lipids, the fatty acid composition was 17.2% and 12.4%, respectively. 115 g of reconstituted lipid in the form of triglycerides were obtained.

Example 2

A solution of 4 g of water mixed with 15 g of glycerine was added to 100 g of a fat or oil containing 42% arachidonic acid derived from Mortierella alpina and stirred for 10 minutes. 50 g of Rhizomucor miehei lipase (Novozymes A / S) was added thereto, and stirred at 200 rpm for 10 hours at 35 ° C. to give arachidonic acid fats containing a large amount of diglycerides and monoglycerides. Prepared. 100 g of 60% gamma linolenic acid (Lipozen, Inc) in the form of free fatty acid derived from borage seed oil was added to the reaction product and stirred under a reduced pressure of 10 ° C. and 10 ⁻² torr for 48 hours to further enhance arachidonic acid. Acid-containing triglyceride reconstituted lipids were prepared. Unreacted free fatty acids and mono- and di-glycerides were removed through molecular distillation to obtain 136 g of a triglyceride composition having arachidonic acid content of 21.0% and gamma linolenic acid content of 31.9%.

Example 3

A solution of 4 g of water mixed with 15 g of glycerin was added to 100 g of borage seed oil containing 20% of gamma-linolenic acid and stirred for 10 minutes. 50 g of Rhizomucor miehei lipase (Novozymes A / S) was added and stirred at 200 rpm for 10 hours at 35 ° C. to obtain borage seed oil containing a large amount of diglyceride and monoglyceride. Prepared. 100 g of a fat or oil in the form of ethyl ester containing 76% arachidonic acid derived from Mortierella alpina was added to the reaction product, and stirred for 60 hours at 60 ° C. under a reduced pressure of 10 ⁻² torr. Enhanced gamma linolenic acid-containing triglyceride reconstituted borage seed oil was prepared. Unreacted ethyl ester and mono- and di-glycerides were removed by molecular distillation to obtain 129 g of a triglyceride composition containing 33.4% arachidonic acid and 15.9% gamma linolenic acid. .

As described above, the food supplement composition of the present invention helps the positive metabolism of arachidonic acid in the application of arachidonic acid to baby food, and at the same time minimizes the possibility of side effects of inflammatory diseases due to excess metabolism of arachidonic acid, saturated fatty acid Functional fatty acid compositions can be provided that minimize other lipids.

Claims (14)

Arachidonic acid-source characterized in that it comprises a reconstituted triglyceride represented by the formula 1, food supplement composition for inhibiting in vivo side effects of arachidonic acid: Formula 1

In the above formula, at least one of R ₁ , R ₂ and R ₃ is arachidonic acid and at least one is gamma-linolenic acid. The food supplement composition of claim 1, wherein the content of the reconstituted triglycerides is 0.1-80% by weight relative to the total composition. 2. The food supplement composition of claim 1 wherein the arachidonic acid in the reconstituted triglyceride is at least 10% and the gamma-linolenic acid is at least 5%. According to claim 1, wherein the reconstituted triglyceride is a single type triglyceride reconstituted microorganism-derived, egg yolk-derived or fish oil-derived arachidonic acid-linked triglycerides and vegetable seed oil-derived gamma-linoleic acid through the enzyme method using lipase Food supplement composition characterized in that. The food supplement composition of claim 1, wherein the composition is a baby food supplement composition. Method for preparing a reconstituted triglyceride represented by the following formula (1) comprising the following steps: Formula 1

In the above formula, at least one of R ₁ , R ₂ and R ₃ is arachidonic acid and at least one is gamma-linolenic acid. (a) hydrolyzing the microorganism-derived, egg yolk-derived or fish oil-derived arachidonic acid-linked triglyceride to obtain arachidonic acid in free arachidonic acid or ethyl ester form; And (b) adding lipase to the mixture of arachidonic acid, gamma-linoleic acid and glycerin in free fatty acid or ethyl ester form and reacting at 50-65 ° C. and 10 ⁻¹ -10 ^-3 torr pressure to bind arachidonic acid and gamma-linoleic acid Obtaining triglycerides. Method for preparing a reconstituted triglyceride represented by the following formula (1) comprising the following steps: Formula 1

In the above formula, at least one of R ₁ , R ₂ and R ₃ is arachidonic acid and at least one is gamma-linolenic acid. (a) a lipase treatment of a mixture comprising (i) microbe-derived, egg yolk-derived or fish oil-derived arachidonic acid-linked triglycerides, (ii) glycerin and (iii) water to mono-glycerides and diglycerides Forming a; And (b) adding gamma-linoleic acid to the reaction product of step (a) and reacting at a temperature of 50-65 ° C. and 10 ⁻¹ -10 ^-3 torr to obtain a triglyceride combined with arachidonic acid and gamma-linoleic acid . Method for preparing a reconstituted triglyceride represented by the following formula (1) comprising the following steps: Formula 1

In the above formula, at least one of R ₁ , R ₂ and R ₃ is arachidonic acid and at least one is gamma-linolenic acid. (a) treating lipase with a mixture comprising (iii) gamma-linoleic acid-linked triglycerides, (ii) glycerin and (iii) water to form mono-glycerides and diglycerides; And (b) adding arachidonic acid in the form of free arachidonic acid or ethyl ester to the reaction product of step (a) and reacting at 50-65 ° C. and 10 ⁻¹ −10 ⁻³ torr pressure to bind arachidonic acid and gamma-linoleic acid Obtaining triglycerides. The method according to any one of claims 6 to 8, wherein the lipase is Candida rugosa , Candida antartica , or Rhizomucor miehei -derived lipase. Way. 9. The process according to claim 6, wherein step (b) is carried out at 60 ° C. temperature and 10 ⁻³ torr pressure. 10. The method according to claim 7 or 8, wherein step (a) is carried out at a temperature of 30-40 ° C. and atmospheric pressure. 8. A process according to claim 6 or 7, wherein the arachidonic acid-linked triglycerides used in step (a) are microorganism-derived. 13. The method of claim 12, wherein said arachidonic acid-binding triglyceride is Mortierella alpina -derived fat or oil. Arachidonic acid-source and food supplement composition for inhibiting in vivo side effects of arachidonic acid, characterized in that it comprises a reconstituted triglyceride prepared by the method of any one of claims 6 to 8.

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