JP5169827B2 - Antiallergic composition and agent, food and drink containing these, and feed - Google Patents

Description

TECHNICAL FIELD OF THE INVENTION

  The present invention is characterized by having β-1,4-mannobiose as a main component, having an inhibitory effect on mast cell degranulation, and having an inhibitory effect on histamine release, which is an important substance for the development of allergy. It is a composition, or food and drink or feed containing them.

Conventional invention

  Recently, allergic diseases have been on the rise, and the causes are clear, such as an increase in Japanese cedar pollen, an increase in allergens due to the living environment, changes in dietary habits due to Western diets, and air pollution. Absent. As allergic diseases, allergic rhinitis, atopic dermatitis, bronchial asthma and the like are the main increasing diseases.

  Allergy is said to occur when the balance of activation of Th1 cells and Th2 cells tilts toward Th2. Th1 cells are responsible for cell immunity and are responsible for T cell proliferation, differentiation and macrophage activation. Th2 cells are referred to as humoral immunity and are responsible for B cell proliferation and differentiation. ing. These B cells produce what is called immunoglobulin E (IgE). If this balance is too inclined to Th2, excess IgE is produced, and it binds to receptors on mast cells, calcium balance is lost, osmotic pressure in mast cells is reduced, and water moves into mast cells. And eventually rupture (degranulate). These mast cells contain substances such as histamine, serotonin, and leukotriene, which are released along with the degranulation of mast cells. In the case of histamine, it binds to the H1 receptor in the body, causing nasal discharge, redness / itchiness.・ It causes allergic symptoms such as edema, pain and bronchoconstriction. IgE originally exhibits an antigen-antibody reaction against parasites and the like and should not be secreted by allergens or pollen in food. For example, when allergens in foods enter the body, the immune system in the body must be ignored by oral tolerance in the intestine. However, if the immune system is tilted to the Th2 side, this oral immune tolerance will not work, and IgE will be produced at random, resulting in allergic symptoms. Therefore, as a countermeasure for allergies, improving the balance of the immune system of Th1 and Th2 is a major issue, but apart from the daily precautions of improving eating habits, You can think like this. 1) Method of forcibly introducing bacteria into the body and activating only Th1 cellular immunity, such as influenza and tuberculin reactions 2) Promoting production of IgA and not allowing antigen to enter the body 3) Chemistry in mast cells Prevent the substance from binding to the H1 receptor 4) Act directly on mast cells to suppress degranulation.

  The method 1) cannot be said to be very safe and the burden on the body is large. It depends greatly on future medical development. Regarding the promotion of IgA production of 2), JP-A-2005-75740 discloses that a mushroom extract promotes production of IgA. It is already on the market in large quantities as a commercially available allergy or cold medicine that does not bind to the H1 receptor of 3). Finally, 4) is a device that works directly on mast cells to suppress degranulation, but this research has just started. For example, J Pharmacol sci 98,90-93 (2005) discloses that prostaglandins have the effect of inhibiting degranulation and inhibiting histamine release by blocking IgE from acting on mast cells. ing. Biol.Pharm.Bull 28 (8) 1438-1443 (2005) discloses that the curcumin compound has a function to prevent degranulation by having some influence on calcium transmission in mast cells. However, this is an advanced compound, and there are economic problems to obtain. Based on the above situation, the present invention is a safe, economical, and naturally occurring component that does not put a burden on the body, and does it work directly on mast cells to suppress degranulation? Was obtained as a result.

  The problem to be solved by the present invention is to provide an antiallergic composition or agent that is excellent in safety, works directly on mast cells to prevent degranulation, and suppresses histamine release, and foods and drinks and feeds containing them. It is to provide.

  As a result of intensive investigations to solve such problems, the present inventors have made mannan degrading enzyme act as a raw material from copra meal and palm kernel meal, hydrolyzed, and removed impurities using ethanol. The present inventors found that a water-soluble component having 10% by weight or more of β-1,4-mannobiose as a main component works directly on mast cells and has a histamine release inhibitory effect, and has led to the present invention. .

  That is, in the present invention, a mannan-degrading enzyme is allowed to act on copra meal and palm kernel meal, and in a water-soluble component obtained by removing contaminants with ethanol, at least 10% by weight or more of β-1,4- more than mannan before decomposition An antiallergic composition having an effect of suppressing degranulation of mast cells, characterized by comprising mannobiose as a main component, and includes a food and drink and a feed containing them.

Hereinafter, the present invention will be described in detail.
In the present invention, β-1,4-mannobiose is formed by two D-mannose molecules linked by β-1,4-glycosides. In the present invention, β-1,4-mannobiose can be obtained by, for example, a method of synthesizing from mannose or a method of decomposing β-1,4-mannan (hereinafter also simply referred to as mannan).

  The method of decomposing β-1,4-mannan is more preferable in terms of the raw material resource and reaction efficiency, and β-1,4-mannobiose can be obtained more easily. In this method, for example, mannan-degrading enzyme is allowed to act on mannan-containing natural products such as coconut cake rich in mannan, palm kernel meal, copra meal, guar gum, locust bean gum, or mannan extracted from these natural products, β-1,4-mannobiose can be obtained.

  Coconut cake refers to a solid material obtained by solid-liquid decomposition of a slurry obtained by crushing endosperm in coconut fruit, and is used for food. Copra meal generally refers to a residue obtained when palm oil is extracted from copra obtained by drying core meat in coconut pulp by sun-drying or hot-air drying, but in the present invention, it does not go through a sun- or hot-air drying step. The extracted oil residue is also included. In addition, the extraction method of palm oil is not specifically limited, such as what extracted using the solvent, the extruder, or the combination of these. Palm kernel meal is a residue obtained by extracting palm kernel oil from palm kernels, which are seeds of oil palm, and can also be extracted by solvent extraction, extruder extraction, or a combination thereof, but is not particularly limited. . Among these, since coconut cake is used for food, it is more preferably used in that the cost can be reduced by omitting the extraction and purification of mannobiose described later.

  In the present invention, the degradation of mannan must be enzymatic hydrolysis. Typical examples of the hydrolysis method include acid decomposition. However, hydrolysis by these methods tends to cause acid modification of the resulting composition, and it is difficult to obtain the expected effect. The enzyme used is not particularly limited as long as it decomposes mannan and produces at least 10% by weight of β-1,4-mannobiose with respect to mannan before decomposition, such as mannanase and mannosidase. Hemicellulase can be used, for example, a commercially available preparation, a culture solution obtained by culturing microbial cells, or a product isolated from microbial cells can be used. As commercially available preparations, for example, hemicellulase GM “Amano” (manufactured by Amano Pharmaceutical Co., Ltd.), Sumiteam ACH (manufactured by Shin Nippon Chemical Industry Co., Ltd.), cellulosin GM5 (Hankyu BioIndustry Co., Ltd.) and the like can be preferably used. In addition to these, even those commercially available as xylanase and cellulase, those having the hydrolytic activity can be used, for example, cellulase Y-NC (manufactured by Yakult Pharmaceutical Co., Ltd.) can be used. In particular, hemicellulase GM “Amano” (manufactured by Amano Pharmaceutical Co., Ltd.) and Sumiteam ACH (manufactured by Shinnippon Chemical Co., Ltd.), which have low mannosidase (exo type) activity and high mannanase (endo type) activity, produce mannose. It is preferable in that it can be suppressed and a large amount of mannobiose can be generated.

  Furthermore, the enzyme used in the present invention acts on mannan-containing natural products or mannan extracted therefrom as an enzyme solution dissolved or dispersed in water. In order to perform an efficient reaction in the case of using a mannan-containing natural product, it is important to adjust moisture in a reaction system including the mannan-containing natural product, a mannan-degrading enzyme, and water. The amount of water added to adjust the water content is preferably 50 to 10,000 parts by weight, more preferably 50 to 1500 parts by weight, with respect to 100 parts by weight of mannan. By setting the amount of water to be in such a range, the mannan fibers can be sufficiently swollen in the presence of sufficient moisture, and the enzyme solution can be easily contacted. However, an excessive amount of water not only decreases the reaction efficiency as a result of diluting the enzyme concentration, but also increases the drying cost in the drying step when drying. Therefore, considering the above, it is appropriate to add 50 to 500 parts by weight of water to 100 parts by weight of mannans.

  The amount of the enzyme and the reaction time are at least 10% by weight of the mannobiose produced with respect to the mannan before decomposition, and preferably the mannobiose produced by hydrolysis is about 10 to 80% by weight with respect to the mannan before decomposition. If it is a thing, it will not specifically limit, Under such conditions, a wet enzyme processed material can be obtained. However, since an enzyme having a high mannanase (endo type) activity usually also has a mannosidase (exo type) activity, if the enzyme reaction time is too long, mannobiose is decomposed and the amount of mannose increases. Therefore, the reaction time is preferably not longer than necessary. These enzyme reaction conditions are appropriately set so that the amount of mannobiose produced is as large as possible. In this case, it is preferable to set so that β-1,4-mannobiose is contained more than mannose. For example, the ratio of mannose to β-1,4-mannobiose is more preferably 60% by weight or less, and 20 It is particularly preferred that it is not more than wt%.

  As described above, for example, when palm kernel meal (mannan content is approximately 36%) is used as a raw material and reacted for 3 to 36 hours, the amount of mannobiose depends on the type, amount, and time of the enzyme used. However, it can be produced up to about 6 to 17 parts by weight with respect to 100 parts by weight of the raw material.

  The obtained decomposition product is preferably dried, but may be an aqueous solution composition as it is. The drying method is not particularly limited, and examples thereof include freeze-drying, spray drying with addition of excipients such as dextrin, fluidized bed drying, and the like. Moreover, ethanol is utilized in order to remove impurities from the obtained enzyme degradation product, if necessary. Other possible solvents that can remove impurities are methanol, isopropanol, hexane, and the like, and ethanol is preferred from the viewpoint of safety. After removing impurities with ethanol, water extraction may be performed. In this case, since β-1,4-mannobiose considered as a main component is water-soluble, a concentration operation may be performed. The drying method of the obtained water-soluble component is not specifically limited. In particular, the aqueous solution composition may be used as it is without being dried. Examples of the drying method include freeze drying, spray drying with addition of excipients such as dextrin, fluidized bed drying, and the like.

  The β-1,4-mannobiose content is preferably hydrolyzed so as to be 10% by weight or more based on the mannan weight before hydrolysis. The mast cell degranulation inhibitory effect expected when this content is less than 10 is not observed. It is not certain whether the mast cell degranulation inhibitory effect seen in the present invention is caused only by this β-1,4-mannobiose, but it is expected to act as at least a main component. However, since it is limited to coconut cake, copra meal and palm kernel meal as mannan-containing compositions in order to obtain a more expected mast cell degranulation inhibitory effect, trace amounts of components and β- It is thought that a unique effect is shown by the synergistic action of 1,4-mannobiose. Therefore, even if the amount of β-1,4-mannobiose is 100%, the effect of the present invention is considered to be obtained. However, it is economically burdensome to extract β-1,4-mannobiose with high purity, In addition, since a synergistic effect with the estimated trace component cannot be expected, the content of β-1,4-mannobiose is preferably 10% by weight or more and less than 90% by weight, more preferably 15% by weight or more and less than 40% by weight.

  The antiallergic composition having an inhibitory effect on degranulation of mast cells of the present invention may be appropriately formulated by a public method and used in the form of an intestinal immunity stimulating agent. , Breads, confectionery, vitamins and other health foods can be added and eaten as foods and drinks, with no particular restrictions. Similarly, it can be used as feed by adding it to fishery and land animal feed.

Hereinafter, the present invention will be described specifically by way of examples. The present invention is not limited to these examples.
<Sample preparation>
Coprameal containing 30% mannan and coprameal A (oiled without going through the drying step) Dried residue), palm kernel meal, or 100 parts by weight of coconut cake, and after 12 hours at 60 ° C., fluidized bed drying was performed until the water content was less than 10% to obtain a dry powder.

  Two times the amount of ethanol was added to 100 parts by weight of the dried product, and the mixture was stirred and then filtered. The filtrate obtained by adding 500 parts by weight of warm water at 60 ° C. and performing filtration after stirring was freeze-dried to obtain the antiallergic composition of the present invention. The sugar composition was as follows.

Next, the following experiment was conducted in order to confirm the mast cell degranulation inhibitory effect according to Examples 1 and 2.
400 μl of Tyrode solution containing 0.05% BSA was added to a microplate containing 400 μl of a suspension of rat mast cells (RBL-2H3) (2 × 10 ⁵ cells / ml), and the concentration was adjusted to 50 μg / ml. Examples 1 and 2 50 μl of an aqueous solution was added, and carbon dioxide culture was performed at 37 ° C. for 1 hour.

Thereafter, 50 μl of 1 μg / ml A23187, which is a calcium transport antibiotic, was added, the reaction was stopped after 5 minutes, the supernatant was collected by centrifugation, and the histamine concentration was measured. The concentration of histamine was measured using a histamine EIA kit (Labor Diagnostika Nord GmbH & Co. KG).
Table 2 shows the inhibitory effect on histamine release in inhibiting degranulation of mast cells in Examples.

  As can be seen from the above results, it can be seen that the product of the present invention acts on mast cells and significantly suppresses the release of histamine, and functions as an antiallergic composition.

  According to the present invention, intestinal immunity activation that can increase IgA production, an antibody important for intestinal immunity, characterized by comprising a plant mannan degradation product containing β-1,4-mannobiose as a main component. It has an action and can be used for a composition having a disease-preventing function due to pathogenic bacteria or viruses, or a food or drink containing them.

Claims (2)

Contains β-1,4-mannobiose obtained by allowing a mannan degrading enzyme to act on coconut cake, copra meal or palm kernel meal so that 10% by weight or more of β-1,4-mannobiose is produced per mannan weight contained An antiallergic agent having a mast cell degranulation inhibitory effect comprising a water-soluble component of the composition as an active ingredient. A β-1,4-mannobiose-containing composition obtained by allowing a mannan degrading enzyme to act on coconut cake, copra meal or palm kernel meal so that 10 wt% or more of β-1,4-mannobiose is produced per mannan weight contained The antiallergic agent according to claim 1, which has an inhibitory effect on mast cell degranulation, comprising as an active ingredient a component extracted by adding water to a residue component extracted by drying the product and adding ethanol.