JP3366770B2 - Antiallergic nutrition composition - Google Patents

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an antiallergic nutritional composition, particularly an antiallergic nutritional composition in which protein and milk-derived fat globule membrane components are treated with an enzyme and blended to reduce the antigenicity of the protein. Regarding things. INDUSTRIAL APPLICABILITY The antiallergic nutritional composition of the present invention is useful for preventing and treating allergic diseases in infants and adults.

[0002]

2. Description of the Related Art Food allergy is an immunological disease caused mainly by proteins, and especially milk,
Proteins derived from eggs or soybeans are heterogeneous proteins for humans, are not sufficiently decomposed in the digestive tract, and are absorbed into the body with antigenicity, and thus are likely to be causative substances of food allergies. Such food allergies are particularly likely to occur in infants with an undeveloped digestive tract mechanism, and their clinical medical conditions are diverse. It is said that removal of allergens from food is the basis for prevention and treatment of food allergies. At present, various nutritional compositions for the treatment or prevention of allergies are commercially available. These commercially available antiallergic nutritional compositions are used as a nitrogen source by hydrolyzing a protein with an enzyme to form peptides or free amino acids. Most of these are mixed.

The production of such an anti-allergic nutritional composition is generally carried out by mixing and dissolving lipids, sugars, vitamins and minerals in addition to the peptide or free amino acid as a nitrogen source, and homogenizing After passing through the sterilization step, it is carried out by pulverizing by concentration and drying. However, when the nitrogen source is incorporated in the form of peptide or free amino acid, the emulsion stability is significantly reduced during the production process, and precipitation or fat separation occurs, unlike the case of incorporation in the form of protein. There is a problem of losing value. Therefore, for the purpose of increasing the emulsion stability, for example, sucrose fatty acid ester, glycerin fatty acid ester, propylene glycol fatty acid ester, sorbitan fatty acid ester and the like chemically synthesized emulsifiers, soybean lecithin, egg yolk lecithin and other natural emulsifiers are used. . In addition, as an example of using a milk fat globule membrane component as an emulsifier, O / W for oral administration
A method for producing a dry powder of a type emulsion is disclosed (JP-A-4-244020), which is used for the purpose of emulsifying a mixture of a fat-soluble vitamin and an oil and fat.

[0004]

The chemical synthetic emulsifier used in the above-mentioned conventional antiallergic nutritional composition is recognized as an additive for general foods, but it is used for the treatment and prevention of allergies. Nutritional composition, especially
Addition of such chemically synthesized emulsifiers to nutritional compositions for allergic diseases targeting food allergies for infants may be unresolved in terms of safety, etc. depending on the type of emulsifier, so it is easy to use. Can not do it. In addition, there is a problem that the flavor is inferior, which is not preferable. Similarly, soybean lecithin and egg yolk lecithin used in the above conventional anti-allergic nutritional composition is a natural emulsifier, but soybean or egg that causes allergies is used as a raw material and extracted from it. Therefore, when added to the antiallergic nutrition composition, it may become a new allergen. In fact, it has been reported that allergies are caused by a protein contained in a small amount in soybean lecithin or egg yolk lecithin. Therefore, soybean lecithin or egg yolk lecithin cannot be used as an emulsifying agent in an antiallergic nutritional composition for people who are likely to develop allergies due to soybeans or egg proteins. Furthermore, the milk fat globule membrane component has little function as an emulsifier in an antiallergic nutritional composition containing a peptide or amino acid obtained by enzymatically treating a protein as a nitrogen source, and is a trace amount. Since it contains milk protein, there is a problem that it becomes a new allergen.

In view of such a situation, the present inventors have found that when the antiallergic nutritional composition is compounded as an emulsifier,
As a result of diligent research on a component having a high emulsifying function, which does not become an allergen, has a good flavor and is safe as a food, it is confirmed that the enzyme-treated milk-derived fat globule membrane component satisfies all the requirements. Heading, completed the present invention. That is, an object of the present invention is to provide an anti-allergic nutrition composition that solves the above-mentioned problems of the prior art.

[0006]

Means for Solving the Problems The present invention, molecular weight of 500
Containing 0 or less peptides and / or free amino acids
Protein so that the molecular weight of the protein
It comprises an anti-allergic nutritional composition containing a milk-derived fat globule membrane component enzymatically treated with ase and peptidase as an active ingredient. The present invention also comprises the above-mentioned anti-allergic nutritional composition containing 0.2% by weight or more of enzyme-treated milk-derived fat globule membrane component per solid content . Further, the present invention is to mix the fat globule membrane component derived from the protein and milk, both
So that the molecular weight of the
And the above-mentioned anti-allergic nutritional composition prepared by enzymatic treatment with peptidase . Further, the present invention is
Protein and milk-derived fat globule membrane components each have a molecular weight of 5
000 or less protease and peptidase
The above-mentioned anti-allergic nutritional composition is prepared by enzymatic treatment with ze .

The present invention will be described in detail below. The antiallergic nutritional composition of the present invention can be used for the treatment and prevention of allergic diseases in infants and adults, especially for dietary treatment. However, it can also be used as a material for meals of patients with all diseases other than allergic patients and healthy people. The antiallergic nutritional composition of the present invention can be used in a liquid form or a powder form obtained by drying the liquid form, but is not limited thereto and can be in any other form.

The antiallergic nutritional composition of the present invention contains a peptide having a molecular weight of 5000 or less and / or a free amino acid as a nitrogen source. The molecular weight of the peptide is 500
The reason why it is set to 0 or less is that if the molecular weight is more than 5,000, the antigenicity of the protein will not be lost, which is not preferable as a nitrogen source for the antiallergic nutrition composition. Further, the molecular weight of the peptide is preferably 200 or more. This is because when the molecular weight is less than 200, free amino acids, particularly hydrophobic amino acids having a strong bitterness, increase and the bitterness becomes stronger. The peptide having a molecular weight of 5000 or less and / or the free amino acid can be obtained by enzymatically treating a protein. Suitable examples of the raw material protein include milk protein, egg protein and soybean protein. As milk protein, skim milk, casein, cheese whey, acid whey, whey protein concentrate (WPC), etc., as egg protein, egg yolk protein, egg white protein, ovalbumin, etc., as soybean protein, defatted soybean protein , Isolated soybean protein, concentrated soybean protein and the like can be used. However, as the raw material protein, other than these, for example,
Proteins such as wheat gluten, fish meat protein, animal meat protein and collagen may be used. Enzymatic treatment of proteins is carried out by a conventional method in which a solution of these proteins is treated with a protease or peptidase at 40 to 50 ° C. for about 5 to 24 hours to hydrolyze the proteins, and peptides and / or free peptides having a molecular weight of 5000 or less are released. It can be performed by changing to an amino acid. The peptide and / or free amino acid having a molecular weight of 5000 or less thus obtained is preferably added to the antiallergic nutritional composition of the present invention as a nitrogen source in an amount of 5 to 30% by weight based on the solid content. . If the blending amount is less than 5% by weight based on the solid content, the protein does not satisfy the prescribed amount even if a predetermined amount of the nutritional composition is ingested, and if it exceeds 30% by weight, the protein intake becomes excessive. Absent.

The antiallergic nutritional composition of the present invention is mainly composed of lipids, sugars, vitamins and minerals in addition to the above-mentioned peptide having a molecular weight of 5000 or less as a nitrogen source and / or free amino acid. However, it may contain any other component that can be incorporated into the nutritional composition, for example, dietary fiber, nucleic acids such as nucleotides, flavors, coloring agents, carnitine and the like. In addition, a drug such as an antiallergic agent may be blended according to the symptoms of the patient. Examples of the sugar include starch, soluble polysaccharides, dextrin, sucrose, lactose, maltose, glucose, oligosaccharides, chitin, chitosan, and artificial sweetener aspartame. It is preferable that 40 to 80% by weight of sugar is blended per solid content of the antiallergic nutritional composition of the present invention. As the lipid, butter, lard, fish oil, palm oil, safflower oil, rapeseed oil, coconut oil and the like oils and fats derived from animals and plants can be mentioned as a preferred example, and to these oils and fats, fractionation, hydrogenation and esters. It may be subjected to a process such as exchange. The lipid is preferably added in an amount of 40% by weight or less based on the solid content of the antiallergic nutritional composition of the present invention. The vitamins may be any kind of vitamins, for example, vitamins A, B, C, D, E,
One or more of Ks, folic acid, pantothenic acid, β-carotene, nicotinic acid amide, inositol, choline, etc. are appropriately selected and blended according to the purpose. Examples of minerals include calcium, magnesium, phosphorus, manganese, potassium, sodium, chlorine, iron, copper, zinc and the like, or salts thereof. It is preferable that the vitamins are mixed in an amount of 10 to 15 mg% by weight and the mineral is also added in an amount of 1 to 10% by weight, based on the solid content of the antiallergic nutritional composition of the present invention.

The antiallergic nutritional composition of the present invention further contains an enzyme-treated milk-derived fat globule membrane component.
The milk-derived fat globule membrane component is a film that coats the fat globule of milk, and it plays a role of maintaining the emulsified state of the highly hydrophobic fat globule and the surrounding aqueous phase by the surfactant action. , Mainly composed of proteins, phospholipids and glycolipids. Here, milk can be exemplified by milk such as cow's milk, goat's milk, sheep's milk and human milk, but milk is particularly preferable. In the present invention, as the milk-derived fat globule coating component, butter serum produced as a by-product when producing butter oil from butter or milk cream, or butter milk as described in JP-A-5-292880. PH 4.4-
It is also possible to use a powder containing a complex lipid obtained by adjusting to 4.6 and removing the precipitate of protein produced by isoelectric focusing, and subjecting the supernatant to ultrafiltration or microfiltration and drying. .

When the antiallergic nutritional composition of the present invention is produced, the protein and the fat globule membrane component derived from milk may be mixed and treated with an enzyme at the same time. You may mix separately, after separately treating with an enzyme. That is, in the present invention, when the peptide and / or amino acid contained in the antiallergic nutritional composition is made of milk protein as a raw material, it is possible to perform enzyme treatment with the same enzyme. The fat globule membrane component of (1) can be mixed and treated with an enzyme at the same time, whereby the manufacturing process can be simplified, which is preferable. On the other hand, when the peptide and / or amino acid is soybean protein or egg protein, the substrate of these protein and the fat globule membrane component derived from milk is different, and therefore each of them is separated by a different enzyme that is compatible with each substrate. After treatment, it is preferable to mix. Thus, when the milk-derived fat globule membrane is solely treated with an enzyme, it can itself be an antiallergic nutritional composition. In other words, the enzyme-treated milk-derived fat globule coating component can be used in the form of an oral nutrition composition in the form of liquid, powder, capsule or granule. The enzyme treatment of the milk-derived fat globule membrane component is preferably carried out so that the molecular weight of the protein contained in the fat globule membrane component is 5000 or less. If the molecular weight of the protein is more than 5,000, it may become a new allergen, which is not preferable. The enzyme treatment is
Molecular weight 5000 used as nitrogen source in the present invention
It can be carried out by a method similar to the following enzymatic treatment of peptide and / or free amino acid. Here, the characteristics of the raw material milk-derived fat globule coating component and the enzyme-treated milk-derived fat globule coating component used in the present invention are shown in Table 1 below.

[0012]

[Table 1] −−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−                               Protein content                               Molecular weight distribution Phospholipid content −−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−− Fat globule film component 20,000-150,000 About 80% Enzyme-treated fat globule membrane component 5,000 or less About 80% −−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−

[0013] The milk-derived fat globule membrane component dramatically improves the emulsifying ability for an anti-allergic nutritional composition, which was hardly possessed before the enzyme treatment, by treating with an enzyme, and the milk-derived fat globule. The milk protein contained in the film component is also decomposed and the antigenicity is significantly reduced. The enzyme-treated milk-derived fat globule membrane component is preferably added to the antiallergic nutritional composition of the present invention in an amount of 0.2% by weight or more based on the solid content. If the blending amount is less than 0.2% by weight, the emulsification is not stable, so that precipitation or fat separation may occur during the production process or during the distribution process when the liquid is distributed. It is preferable to secure the blending amount. The antiallergic nutritional composition of the present invention is a mixture of the above-mentioned components and a milk-derived fat globule membrane component as an emulsifier, which is emulsified by a conventional method to obtain a homogeneous
It is possible to fill the container as a liquid through a sterilization step or, if necessary, further to a powder through a concentration / drying step.

Hereinafter, the present invention will be described in detail with reference to Examples, and Comparative Examples and Test Examples will be shown to confirm the effects of the present invention.

【Example】

Example 1 (Preparation of milk-derived fat globule coating component) 10 kg of butter 75
Dissolve at ℃, add 2 kg of warm water at the same temperature, stir well, then centrifuge with a continuous centrifuge (5,000 x
g) and fractionated into butter oil and butter serum.
2 kg of the fraction collected as butter serum was freeze-dried,
140 g of fat globule coating component was obtained. (Preparation of antiallergic nutritional composition) 100 g of the fat globule membrane component obtained above and 7 kg of casein powder were dissolved in 93 kg of water and the pH was adjusted to 3.9, and 560,000 units of protease (Amano Pharmaceutical Co., Ltd. ); Protease M) was added and treated at 46 ° C. for 6 hours with stirring. Furthermore, after adjusting the pH of this solution to 6.1, 30 million units of peptidase (manufactured by Amano Pharmaceutical Co., Ltd .; Peptidase R) were added and treated at 46 ° C. for 16 hours while stirring. 100 kg of the obtained hydrolyzed liquid, 6 kg of purified tapioca starch, 30 kg of dextrin, 2.5 kg of sucrose,
Predetermined amount of water-soluble vitamins (B ₁ , B ₂ , B ₆ , B ₁₂ ,
C, niacin, folic acid, pantothenic acid), and a predetermined amount of salts (sodium citrate, calcium carbonate, magnesium chloride, sodium ferrous citrate, copper sulfate, zinc sulfate) were added and dissolved while heating. In this solution,
A predetermined amount of fat-soluble vitamins (A, D, E,
5 kg of safflower oil mixed with K) was added and homogenized, and then spray-dried to prepare an antiallergic nutritional composition (A) 5
I got 0 kg.

Example 2 (Enzymatic treatment of milk-derived fat globule membrane component) 100 g of fat globule membrane component obtained by the same method as shown in Example 1.
Was dissolved in 1 L of water and the pH was adjusted to 3.9, then 5,000
0 unit of protease (manufactured by Amano Pharmaceutical Co., Ltd .; Protease M) was added and treated at 46 ° C. for 6 hours with stirring. Furthermore, after adjusting the pH of this solution to 6.1, 300,000 units of peptidase (manufactured by Amano Pharmaceutical Co., Ltd .; Peptidase R) were added and treated at 46 ° C. for 16 hours while stirring. (Preparation of antiallergic nutritional composition) 7 kg of casein powder was dissolved in 93 kg of water, and after adjusting the pH to 3.9, 560,000 units of protease (Amano Pharmaceutical Co., Ltd .; Protease M) was added to give 46 Treated at 6 ° C with stirring for 6 hours. Furthermore, after adjusting the pH of this solution to 6.1,
30 million units of peptidase (manufactured by Amano Pharmaceutical Co., Ltd .;
Peptidase R) was added and treated at 46 ° C. with stirring for 16 hours. In 100 kg of this hydrolysis treatment liquid,
1 kg of fat globule membrane component solution obtained by the above enzyme treatment, 6 kg of purified tapioca starch, 30 kg of dextrin, 2.5 k of sucrose
g, a certain amount of water-soluble vitamins (B ₁ , B ₂ , B ₆ , B ₁₂ ,
C, niacin, folic acid, pantothenic acid) and a predetermined amount of salts (sodium citrate, calcium carbonate, magnesium chloride, sodium ferrous citrate, copper sulfate, zinc sulfate) were added and dissolved while heating. Add a predetermined amount of fat-soluble vitamins (A, D, E, K) to this solution in advance.
5 kg of safflower oil mixed with was homogenized and then spray-dried to obtain 50 kg of antiallergic nutritional composition (B).

Example 3 7 kg of purified soybean protein powder was dissolved in 93 kg of water, the pH was adjusted to 6.8, and 30 million units of papain (manufactured by Wako Pure Chemical Industries, Ltd.) was added to 60 Treated at 8 ° C. for 8 hours with stirring. 100 kg of this hydrolyzed solution was treated with the enzyme in the same manner as in Example 2 to obtain 1 g of a fat globule membrane component solution, 6 kg of purified tapioca starch, 30 kg of dextrin,
2.5 kg sucrose, a certain amount of water-soluble vitamins (B ₁ ,
_{B 2, B 6, B 12} , C, niacin, folic acid, pantothenic acid) and a predetermined amount of salt (sodium citrate, calcium carbonate, magnesium chloride, sodium ferrous citrate, copper sulfate, sulfuric acid was added zinc) , Dissolved while heating. To this solution, 5 kg of safflower oil mixed with a predetermined amount of fat-soluble vitamins (A, D, E, K) was added and homogenized, and then spray-dried to obtain 50 kg of an antiallergic nutritional composition. .

Example 4 A milk-derived fat globule film component treated with an enzyme was pulverized by the same method as that described in Example 2, and 1 g per tablet was filled in a soft capsule made of gelatin to obtain an antiallergic nutritional composition. .

Comparative Example 1 50 kg of an antiallergic nutritional composition (C) was obtained in the same manner as in Example 2 except that 100 g of soybean lecithin was added instead of the enzyme-treated milk-derived fat globule membrane component. It was

Comparative Example 2 Antiallergic substance was prepared in the same manner as in Example 2 except that 100 g of the non-enzymatic milk-derived fat globule membrane component was added in place of the enzyme-treated milk-derived fat globule membrane component. 50 kg of nutritional composition (D) was obtained.

Test Example 1 (Preparation of antiserum) The nutritional compositions prepared in Examples 1 and 2 and Comparative Examples 1 and 2 were respectively added to 10 mM phosphate buffer (pH 7.2) containing 150 mM Nacl at 20 mg / mg. It was dissolved at a concentration of ml to prepare samples (A), (B), (C) and (D). Emulsified by mixing each sample with 20 mg / ml concentration of aluminum hydroxide solution, and emulsifying each
0.5 ml per mouse was intraperitoneally administered to lb / c mice (5 weeks old, male). After four doses every other week, 4 days after the final dose, blood was collected from the heart to separate serum,
The respective sera were designated as sera (A), (B), (C) and (D).

(Evaluation of Allergenicity) Whether or not the nutritional compositions produced in Examples 1 and 2 and Comparative Examples 1 and 2 have allergenicity was examined by the PCA method. The above serum (A)
(D) is 2 times, 20 times, 2 times with physiological saline, respectively.
50 μl of each was diluted to 00 times and 2,000 times, and 50 μl was intradermally injected into the dorsal skin of SD rats (10-week-old, female) whose back hair was shaved. 24 hours after administration, the samples (A) to
(D) 1 ml of each was mixed with 9 ml of a 0.6% Evans blue solution adjusted with physiological saline, and 1 ml of each obtained solution was administered to the rat through the tail vein. Furthermore, after 60 minutes, the rat was sacrificed, and the minor and major diameters of locus ceruleus observed at the serum-administered site on the dorsal endothelium were measured. If the average value was 5 mm or less, it was negative (-). It was defined as positive (+). As a result, the allergenicity of the samples (A) and (B) was negative, but the samples (C) and (D) were positive.

Test Example 2 (Identification of allergen) In each serum prepared in Test Example 1,
The presence or absence of antibodies to the milk or soybean-derived allergens was examined by ELISA. 50 μl each of milk casein solution and soybean protein solution having a concentration of 1 mg / ml were dispensed into a 96-well microtiter plate, and 37
Incubated at 0 ° C for 1 hour. After removing the solution and blocking the plate with a 1% ovalbumin solution, mouse serums (A) to (D) diluted in 20, 200 and 2,000 times prepared in Test Example 1 were dispensed in 50 μl portions.
After incubating at 37 ° C. for 1 hour, peroxidase-labeled anti-mouse immunoglobulin serum (manufactured by Nordic) diluted 200 times was dispensed in 50 μl portions, and
Incubated at 7 ° C for 1 hour. Finally, AB
The TS substrate solution was added to develop color, and the absorbance at 595 nm was measured. As a result, antibodies that react with casein and soybean protein were not detected in the mouse sera (A) and (B) prepared in Test Example 1, but in the serum (C),
An antibody that reacts with soybean protein was detected, and an antibody against casein was detected in serum (D). Therefore, test example 1
The allergens of the nutritional compositions (C) and (D) shown in
It was found that they were caused by soybean protein and casein, respectively.

Test Example 3 (Emulsifying property of milk-derived fat globule coating component treated with enzyme) In order to compare the emulsified states of the nutritional compositions (A) and (D) produced in Example 1 and Comparative Example 2, Each powder 7
After dissolving g in 50 ml of hot water at 50 ° C, 5 at 800 × g
Centrifugation was carried out for a minute to separate into oil phase / emulsion phase / water phase, and the height of each phase was measured and compared. The results are shown in Table 2.

[0024]

[Table 2]                                                           Unit (cm) −−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−                       Height of water phase Height of emulsified phase Height of oil phase −−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−− Nutrition composition (A) 0 7.3 ± 0.5 0.7 ± 0.4 Nutrition composition (D) 1.0 ± 0.2 5.0 ± 0.6 1.1 ± 0.3 −−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−−

As is clear from the results shown in Table 2,
In the nutritional composition (A) using the milk-derived fat globule coating component enzyme-treated as an emulsifier, the oil phase and the water phase were almost emulsified and the emulsified state was stable. On the other hand, in the nutritional composition (D) using the milk-derived fat globule membrane component which was not treated with the enzyme, both the water phase and the oil phase had a part which was not emulsified and the emulsified state was unstable.

[0026]

INDUSTRIAL APPLICABILITY The antiallergic nutritional composition of the present invention comprises an enzyme-treated milk-derived fat globule membrane component as an active ingredient, and a peptide having a molecular weight of 5000 or less and / or a free amino acid as a nitrogen source. Since the above-mentioned enzyme-treated milk-derived fat globule film component is contained as an emulsifier, the antigenicity of the entire composition is extremely low and the emulsion stability is excellent. That is, the anti-allergic nutritional composition of the present invention contains a milk-derived fat globule membrane component treated with an enzyme as an emulsifier, as compared with the case where a milk-derived fat globule membrane component not treated with an enzyme is used. At the same time that the emulsifying function is remarkably improved, a trace amount of milk protein contained in the fat globule membrane component is decomposed, and the allergen is almost disappeared. Furthermore, since the antiallergic nutritional composition of the present invention uses a milk-derived fat globule film component as an emulsifier, it is highly safe as a food, as compared with a nutritional composition using a conventional chemically synthesized emulsifier. . Therefore,
The antiallergic nutritional composition of the present invention has antiallergic properties,
Since it has excellent emulsion stability and safety, it is extremely useful as a nutritional composition for the prevention and treatment of allergic diseases in infants and adults.

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Claims (4)

(57) [Claims] 1. A peptide having a molecular weight of 5000 or less and / or
Or the molecular weight of free amino acid and the protein contained is 50
Protease and peptidase so as to be 00 or less
Antiallergic nutritional composition comprising, as an active ingredient, and fat globule membrane component derived from milk enzyme treated with. 2. A fat globule membrane component derived from milk treated with enzyme, antiallergic nutritional composition according to claim 1, containing more than 0.2% by weight per solid content. 3. A protein and a fat globule membrane component derived from milk are mixed so that the molecular weights of both proteins are 5000 or less.
The anti-allergic nutritional composition according to claim 1 or 2, which is prepared by enzymatic treatment with ase and peptidase . 4. A protein and a milk-derived fat globule membrane component, each having a molecular weight of 5000 or less, are proteases.
The anti-allergic nutritional composition according to claim 1 or 2, which is prepared by enzymatically treating with peptidase .