JP7585360B2 - Foods containing nattokinase - Google Patents

Description

The present invention relates to a food product that contains nattokinase and gelatin, yet is capable of suppressing the decomposition of gelatin during storage.

Thrombosis is a pathological condition in which blood clots form in blood vessels and block blood flow in the circulatory system, and is known to be a factor in the development of serious diseases such as cerebral infarction, myocardial infarction, and pulmonary infarction. Conventionally, methods of preventing the formation of blood clots or dissolving blood clots by administering drugs such as antiplatelet agents, anticoagulants, and thrombolytic agents have been used to prevent or treat thrombosis. However, such drugs are accompanied by side effects and require administration under the supervision of a doctor, and therefore cannot be taken easily or on a daily basis.

On the other hand, since it was reported that natto, one of Japan's traditional foods, contains nattokinase, which has thrombolytic activity, the value of natto as a health food has been reevaluated. However, natto has a unique odor and stickiness, and the reality is that there are many people who do not eat natto. Therefore, in order to make nattokinase easier to ingest, food formulations such as capsules and tablets containing nattokinase have been developed. Such foods containing nattokinase are useful for self-medication because they can be ingested easily and daily without the need for medical supervision.

Foods containing nattokinase are often in the form of tablets or capsules coated with a gelatin film for ease of ingestion, etc. (see, for example, Patent Document 1, etc.). Foods containing nattokinase may be made into a gel by adding gelatin as a gelling agent, or liquid foods containing nattokinase may be made viscous by adding gelatin as a thickening agent.

On the other hand, because nattokinase has a proteolytic effect, foods containing nattokinase and gelatin have the disadvantage that the gelatin is decomposed by nattokinase during storage. For example, in the case of tablets or capsules coated with a gelatin film, such gelatin decomposition can cause the gelatin film to crack and the contents to leak, significantly reducing the commercial value of the product. However, to date, no sufficient research has been conducted into technologies that can suppress the decomposition of gelatin during storage in foods containing nattokinase and gelatin.

Special Publication No. 2013-521262

The object of the present invention is to provide a food product that contains nattokinase and gelatin, while suppressing the decomposition of gelatin during storage.

The inventors conducted extensive research to solve the above problems and discovered that the decomposition of gelatin can be suppressed in foods containing nattokinase and gelatin by adding at least one fat-soluble vitamin selected from the group consisting of vitamins A, E, and K. The present invention was completed through further research based on this knowledge.

That is, the present invention provides the following aspects.
Item 1. A food comprising (A) nattokinase, (B) gelatin, and (C) at least one fat-soluble vitamin selected from the group consisting of vitamins A, vitamin E, and vitamin K.
Item 2. The food according to Item 1, wherein the component (C) is at least one selected from the group consisting of retinol, tocopherol, menaquinone, and derivatives thereof.
Item 3. The food according to Item 1 or 2, which is a capsule comprising a capsule shell formed from the component (B).
Item 4. A method for inhibiting decomposition of gelatin in a food containing nattokinase and gelatin, comprising:
A method for inhibiting the decomposition of gelatin, comprising blending (A) nattokinase, (B) gelatin, and (C) at least one fat-soluble vitamin selected from the group consisting of vitamins A, vitamin E, and vitamin K into a food.

According to the food of the present invention, even if it contains nattokinase and gelatin, the decomposition of gelatin can be suppressed, so the function performed by gelatin in the food can be stably maintained. Furthermore, when the food of the present invention is a capsule or tablet containing gelatin as a capsule membrane or coating membrane, the shape of the gelatin membrane can be stably maintained even if it is stored in a hygroscopic state after opening, and cracking of the gelatin membrane and leakage of contents due to storage can be suppressed.

1. Food The food of the present invention is characterized by containing (A) nattokinase, (B) gelatin, and (C) at least one vitamin selected from the group consisting of vitamin A, vitamin E, and vitamin K. The food of the present invention will be described in detail below.

[(A) Nattokinase]
The food of the present invention contains nattokinase (sometimes referred to as component (A)). Nattokinase is an enzyme that has a fibrinolytic effect and is produced by Bacillus subtilis natto.

The nattokinase used in the present invention can be obtained by a known production method. Specific methods for producing nattokinase include a method of culturing Bacillus subtilis natto, a method of obtaining it from a transformant incorporating a gene encoding nattokinase, and a method of synthesizing it by chemical synthesis. The nattokinase used in the present invention may be obtained by any production method, but from the viewpoint of reducing production costs, etc., it is preferable to use a method obtained by culturing Bacillus subtilis natto.

The nattokinase used in the present invention may be a purified product, but may also be in an unpurified state, so long as it can be incorporated into foods. For example, when using nattokinase obtained by culturing Bacillus subtilis natto, it may be an extract of the culture of Bacillus subtilis natto. Furthermore, it may be nattokinase obtained by subjecting a culture of Bacillus subtilis natto to ion exchange chromatography, gel filtration chromatography, hydrophobic chromatography, or the like to purify the nattokinase, or it may be nattokinase obtained by subjecting a culture of Bacillus subtilis natto to a crude purification process such as solid-liquid separation as necessary, after which water is removed or the culture is dried.

Nattokinase is also commercially available as a powder containing excipients, a crude product, a refined product, etc., and these commercially available products can also be used as nattokinase in the present invention.

In the food of the present invention, the content of component (A) may be appropriately set depending on the form of the food, etc., but may be, for example, 100 FU/g or more, preferably 200 to 10,500 FU/g, and more preferably 300 to 5,000 FU/g.

In the present invention, the "FU" indicating the activity of nattokinase is a fibrinolytic activity unit according to the standard for natto culture extract foods announced by the Japan Health and Nutrition Food Association on January 15, 2003.

[(B) Gelatin]
The food of the present invention contains gelatin (sometimes referred to as component (B)). Gelatin is extracted by applying heat to collagen, which is the main component of animal skin, bones, tendons, etc.
In the present invention, gelatin obtained by a known production method can be used, for example, gelatin produced by heat extraction of crude collagen obtained by treating the skin, bone, tendon, etc. of cows, pigs, chickens, fish, etc. with an acid or alkali. Furthermore, the gelatin used in the present invention may be a hydrolysate, an oxygen decomposition product, a gelatin derivative (e.g., acylated gelatin, etc.), etc.

In the food of the present invention, gelatin can be included for various purposes and in various forms depending on the form of the food. Specifically, in the food of the present invention, gelatin can be included as a capsule coating in the case of a capsule formulation, a coating coating in the case of a coating preparation, a gelling agent in the case of a gel food, a thickener in the case of a liquid food, etc.

In the food of the present invention, the content of component (B) may be appropriately set depending on the intended use of the gelatin, the form of the food, etc., but may be, for example, 3 to 80% by weight, preferably 5 to 50% by weight, and more preferably 10 to 40% by weight.

In addition, in the food of the present invention, the ratio of component (A) to component (B) may be within the range according to the respective contents described above, but for example, the ratio may be 100 to 50,000 FU, and preferably 3,000 to 30,000 FU, of component (A) per 1 g of component (B).

[(C) Fat-soluble vitamins]
The food of the present invention contains at least one fat-soluble vitamin (sometimes referred to as component (C)) selected from the group consisting of vitamins A, vitamin E, and vitamin K. By containing such a specific fat-soluble vitamin, it becomes possible to inhibit the decomposition of gelatin by nattokinase.

The vitamins A refer to vitamin A ₁ (retinol, retinal, retinoic acid), vitamin A ₂ (3-dehydroretinol, 3-dehydroretinal, 3-dehydroretinoic acid), and derivatives thereof. The type of vitamin A used in the present invention is not particularly limited, as long as it can be incorporated into foods. For example, derivatives of vitamin A ₁ and A ₂ include retinol organic acid esters such as retinol acetate, retinol butyrate, retinol propionate, retinol palmitate, retinol linoleate, retinol octylate, retinol laurate, retinol oleate, and retinol linoleate. These vitamins A may be used alone or in combination of two or more.

Among vitamin A, from the viewpoint of more effectively suppressing the decomposition of gelatin, preferred are retinol and its derivatives, more preferred are retinol organic acid esters, and particularly preferred are retinol acetate.

Vitamin E refers to vitamin E (tocopherol, tocotrienol) and its derivatives. The type of vitamin E used in the present invention is not particularly limited, as long as it can be incorporated into food. For example, tocopherol and tocotrienol may be any of α-, β-, γ-, and δ-, and may be either d- or dl-isomer. In addition, examples of derivatives of vitamin E include tocopherol organic acid esters such as tocopherol acetate, tocopherol succinate, tocopherol linoleate, tocopherol linolenate, tocopherol nicotinate, and tocopherol (linoleate/oleate). These vitamin E may be used alone or in combination of two or more types.

Among vitamin E, from the viewpoint of more effectively suppressing the decomposition of gelatin, tocopherol and its derivatives are preferred, tocopherol organic acid esters are more preferred, and tocopherol acetate is particularly preferred.

The vitamin Ks refer to vitamin K and derivatives thereof, such as vitamin K ₁ (phylloquinone), vitamin K ₂ (menaquinone), vitamin K ₃ (menadione), vitamin K ₄ (menadiol), and vitamin K 5 (4-amino-2-methyl-1-naphthol). The type of vitamin Ks used in the present invention is not particularly limited, so long as they can be incorporated into foods. For example, menaquinone may be any of menaquinone-4, menaquinone-5, menaquinone-6, menaquinone-7, menaquinone-8, and menaquinone-9. In addition, for example, derivatives of vitamin K ₁ and K ₂ include alkylated products, epoxidized products, reduced products, and the like.

Among the vitamin Ks, vitamin K ₂ (menaquinone) is preferred from the viewpoint of more effectively inhibiting the decomposition of gelatin.

In the food of the present invention, one vitamin may be selected from vitamins A, E, and K and used alone, or two or more vitamins may be used in combination.

The content of component (C) in the food of the present invention may be appropriately set depending on the type of component (C) used, the content of gelatin, the form of the food, etc., and may be, for example, 0.001 to 50% by weight, preferably 0.03 to 25% by weight. More specifically, the content of each type of component (C) in the food of the present invention is exemplified by the following ranges.
When component (C) is vitamin A : the vitamin A content is preferably 5 to 50% by weight, more preferably 10 to 30% by weight, and particularly preferably 15 to 25% by weight.
When component (C) is vitamin E : the vitamin E content is preferably 2.5 to 50% by weight, more preferably 5 to 30% by weight, and particularly preferably 7.5 to 20% by weight.
When component (C) is a vitamin K compound : the vitamin K compound is preferably 0.001 to 50% by weight, more preferably 0.01 to 30% by weight, and particularly preferably 0.03 to 20% by weight.

In the food of the present invention, the ratio of component (A) to component (C) may be within the range according to the respective contents described above, and may be, for example, 0.0125 to 625 mg, preferably 0.375 to 312.5 mg, of component (C) per 1000 FU of component (A). More specifically, the ratio of component (A) to component (C) for each type of component (C) in the food of the present invention may be in the following ranges:
When the component (C) is vitamin A : the amount of vitamin A per 1000 FU of the component (A) is preferably 62.5 to 625 mg, more preferably 125 to 375 mg, and particularly preferably 187.5 to 312.5 mg.
When the component (C) is vitamin E , the amount of vitamin E per 1000 FU of the component (A) is preferably 31.25 to 625 mg, more preferably 62.5 to 375 mg, and particularly preferably 93.75 to 250 mg.
When the component (C) is a vitamin K type compound , the amount of vitamin K type compound per 1000 FU of the component (A) is preferably 0.0125 to 625 mg, more preferably 0.125 to 375 mg, and particularly preferably 0.375 to 250 mg.

[Other ingredients]
In addition to the above-mentioned components, the food of the present invention may contain other nutritional components or functional components as necessary. Such nutritional components and functional components are not particularly limited as long as they are usable in food, but may include, for example, vitamins such as vitamin B, vitamin C, vitamin P, niacin, pantothenic acid, biotin, and lycopene; catechin, theaflavin, leucoanthocyanidin, pelargonidin, cyanidin, petunidin, peonidin, petunidin, delphinidin, malvidin, flavone, apigenin, luteonin, apigeninidin, luteionidin, baicalein, quercetin, kaempferol, myricetin, narizin, hesperidin, liquiritigenin, and alpinone. Polyphenols such as taxifolin, genistein, daidzein, daidzin, glycitein, equol, biochanin A, coumestrol, puerarin, formononetin, carthamin, proretin, gallic acid, phenolic acid, chlorogenic acid, ellagic acid, lignan, sesamin, pinoresinol, lariciresinol, secoisolariciresinol, matairesinol, curcumin, coumarin, resveratrol, and hydrolyzed tannins; stomachics such as betaine hydrochloride, carnitine chloride, and bethanechol chloride; minerals such as calcium, sulfur, magnesium, zinc, selenium, and iron; soy protein, eggs Proteins and peptides such as white powder, whey protein, and sardine peptides; amino acids such as glycine, alanine, arginine, aspartic acid, cystine, phenylalanine, taurine, and tryptophan; fatty acids such as linoleic acid, gamma-linolenic acid, and alpha-linolenic acid; caramel color, gardenia color, anthocyanin color, annatto color, paprika color, safflower color, red yeast rice color, flavonoid color, cochineal color, amaranth, erythrosine, allura red AC, new coccine, phloxine, rose bengal, acid red, tartrazine, sunset yellow FCF, and fast green F. Examples of the ingredients include pigments such as CF, brilliant blue FCF, and indigo carmine; flavorings such as flavors and essences of various fruits; seasonings such as citric acid and its salts, malic acid and its salts, tartaric acid and its salts, acetic acid and its salts, lactic acid and its salts, salt, glutamic acid and its salts, mirin, vinegar, natural fruit juice, plant extracts, and cut or powdered fruits and seafood; mushrooms or their extracts such as agaricus, shiitake mushroom extract, lychee mushroom, and yamabushitake mushroom; and other functional materials such as dietary fiber, royal jelly, propolis, honey, chondroitin sulfate, glucosamine, ceramide, and hyaluronic acid. These ingredients may be used alone or in combination of two or more. The content of these ingredients is appropriately set according to the type of ingredients used and the form of the food.

In addition, the food of the present invention may contain bases and additives, etc., as necessary, in order to prepare it into a desired form. Such bases and additives are not particularly limited, so long as they are usable in foods, and examples thereof include water, oils and fats, waxes, hydrocarbons, fatty acids, higher alcohols, esters, water-soluble polymers, surfactants, metal soaps, lower alcohols, polyhydric alcohols, pH adjusters, buffers, antioxidants, UV protection agents, preservatives, flavors, powders, thickeners, dyes, chelating agents, etc. These bases and additives may be used alone or in combination of two or more types. The content of these bases and additives is appropriately set depending on the type of base or additive used, the form of the food, etc.

[form]
The food product of the present invention may be in solid, semi-solid, or liquid form.

The food of the present invention may take the form of a general food or drink, as well as a specific health food, a nutritional supplement, a functional food, or a food for the sick. Specific examples of the food of the present invention may take the form of supplements such as capsules (soft capsules, hard capsules), tablets, granules, powders, or jellies; beverages such as nutritional drinks, fruit juice drinks, or carbonated drinks; or luxury items such as dumplings, ice cream, sherbet, gummies, or candies. Among these foods and drinks, supplements are preferred, capsules, tablets, or granules are more preferred, and capsules are particularly preferred.

When the food of the present invention is a capsule, it is preferable that the (B) component is contained as a capsule shell, and the (A) and (C) components are contained in the contents. When the food is a tablet or granule, the (A) to (C) components may be contained in a mixed state in the tablet or granule, or the food may be a coating preparation in which the (B) component is coated as a coating film on a tablet or granule containing the (A) and (C) components.

2. Method for inhibiting the decomposition of gelatin The method for inhibiting the decomposition of gelatin in a food containing nattokinase and gelatin according to the present invention is characterized in that it comprises blending, together with (A) nattokinase and (B) gelatin, (C) at least one fat-soluble vitamin selected from the group consisting of vitamins A, vitamins E, and vitamins K, into the food.

In the decomposition inhibition method of the present invention, the types and amounts of nattokinase, gelatin, and fat-soluble vitamins used, other ingredients that can be added to the food, and the form of the food are as described in the section "1. Food" above.

The present invention will be described in detail below with reference to examples, but the present invention is not limited to these examples.

In the test examples and formulation examples shown below, the sources, compositions, etc. of the ingredients used are as follows:
Nattokinase powder : Product name "Natto bacteria culture extract NSK-SD40", manufactured by Japan Bioscience Laboratory Co., Ltd.
Retinol acetate : Product name: "Dry Vitamin A Acetate 325 CWS/A", manufactured by DSM Nutrition Japan Co., Ltd.
Tocopherol acetate : dl-α-tocopherol acetate, product name "Dry Vitamin E 50% CWS/A", manufactured by DSM Nutrition Japan Co., Ltd.
Vitamin K2 _: Trade name "Vitamin K2 Kyowa", manufactured by Kyowa Hakko Bio Co., Ltd.
Arginine : Product name "L-arginine", manufactured by Kyowa Hakko Bio Co., Ltd.
Lysine : Product name "Food Additive L-Lysine Hydrochloride", manufactured by Ajinomoto Co., Inc.
Citrulline : Product name "Fermented Citrulline Kyowa", manufactured by Kyowa Hakko Bio Co., Ltd.
Resistant dextrin : product name "Fibersol 2AG", manufactured by Matsutani Chemical Industry Co., Ltd.
Rutin : Product name "αG Rutin P", manufactured by Toyo Sugar Co., Ltd.

Test Example 1
Gelatin plates were prepared and the decomposition effect of nattokinase on the gelatin plates was evaluated in the presence of various components. The specific test methods and results are as follows:

1. Test method
1-1. Preparation of gelatin plate First, purified water was heated to 50°C, and gelatin was added to 20% by weight and methylparaben (preservative) was added to 0.2% by weight, and mixed to prepare a gelatin solution. 10 g of the obtained gelatin solution was spread in a petri dish (inner diameter 9 cm), and the thickness of the gelatin in the petri dish was made uniform. After that, the solution was cooled at 4°C for 1 hour or more to gel the gelatin and form a gelatin plate in the petri dish.

1-2. Gelatin Plate Decomposition Test Test solutions with the compositions shown in Table 1 were prepared, and 0.5 ml of each test solution was dropped onto a gelatin plate in a petri dish, and the dish was left to stand overnight at room temperature with a lid on. After that, the appearance of the gelatin plate was visually confirmed, and the gelatin decomposition resistance was evaluated according to the following criteria.
<Criteria for determining gelatin decomposition resistance>
1: The gelatin plate is slightly dissolved.
2: The gelatin plate has dissolved slightly, and the difference from the part where no drops were applied is clear.
3: About half the thickness of the gelatin plate is dissolved.
4: Most of the gelatin plate has dissolved, with part of the bottom of the dish exposed.
5: The gelatin plate is completely dissolved and the bottom of the dish is completely exposed.

2. Test results The results obtained are shown in Table 1. When a test solution containing only nattokinase was used, the gelatin plate was completely decomposed (Comparative Example 1). Furthermore, even when a test solution containing arginine, lysine, citrulline, indigestible dextrin, or rutin together with nattokinase was used, the decomposition of the gelatin plate could not be suppressed at all (Comparative Examples 2 to 6). In contrast, when a test solution containing retinol acetate, tocopherol acetate, or vitamin _K2 together with nattokinase was used, the decomposition of the gelatin plate was sufficiently suppressed (Examples 1 to 3).

Formulation Examples 1 to 12: Soft Capsules Using the liquid content having the composition shown in Table 2 and the capsule shell having the composition shown in Table 3, soft capsules (containing 300 mg of liquid content and 150 mg of shell per capsule) were produced.

Formulation Examples 13 to 15: Tablets Tablets (700 mg per tablet) having the composition shown in Table 4 were produced.

Formulation Examples 16 to 18: Granules Granules having the compositions shown in Table 5 were produced.

Claims (4)

(A) nattokinase, (B) gelatin, and (C) at least one fat-soluble vitamin selected from the group consisting of vitamins A, E, and K;
The nattokinase contains 100 FU/g or more,
When vitamin E is included as the fat-soluble vitamin, 31.25 to 625 mg of vitamin E is contained per 1000 FU of nattokinase,
When vitamin K is contained as the fat-soluble vitamin, the food contains 0.375 to 250 mg of vitamin K per 1000 FU of nattokinase . The food according to claim 1, wherein the component (C) is at least one selected from the group consisting of retinol, tocopherol, menaquinone, and derivatives thereof. The food product according to claim 1 or 2, which is a capsule containing a capsule shell formed from the component (B). A method for inhibiting decomposition of gelatin in a food product containing nattokinase and gelatin, comprising:
The method includes a step of blending (A) nattokinase and (B) gelatin together with (C) at least one fat-soluble vitamin selected from the group consisting of vitamins A, vitamin E, and vitamin K, into a food product;
The nattokinase is blended in a food product at a concentration of 100 FU/g or more,
When vitamin E is used as the fat-soluble vitamin, 31.25 to 625 mg of vitamin E is used per 1000 FU of nattokinase.
When vitamin K is blended as the fat-soluble vitamin, 0.375 to 250 mg of vitamin K is blended per 1000 FU of nattokinase .