JP5107545B2 - Peyer's board formation inducing composition - Google Patents

Description

  The present invention relates to a Peyer's board formation-inducing composition containing hemicellulose and / or a partially decomposed product thereof as an active ingredient.

  Living organisms, including humans, are constantly exposed to foreign antigens such as microorganisms, or endogenous foreign self-substances such as tumor cells, and are used to protect against various diseases caused by these. It has an immune system as a mechanism.

  However, it is considered that the immune system is reduced in function due to aging, stress, fatigue, irregular life, various environmental factors, and the like, thereby causing various diseases and infectious diseases. Advancing aging society and busy modern society are in a state where there are countless factors that cause a decline in immunity, but strengthening the immunity, which is the defense function of the living body, It is important to maintain the homeostasis.

  Therefore, immunostimulatory substances that activate the immune system have a wide range of uses as foods and medicines. For example, in Patent Document 1 below, hemicellulose obtained from corn hulls and / or partial degradation products thereof are used as active ingredients. An immunostimulant is disclosed. The immunostimulant disclosed in this publication has a cytokine-producing ability promoting effect on ConA stimulation of spleen cells, and activates the systemic immune system via the circulatory system such as blood and lymph. .

  On the other hand, since pathogenic microorganisms that cause diseases mainly enter the host through mucosal surfaces such as the digestive tract and respiratory organs, and infection is established, for example, as disclosed in Non-Patent Document 1 below, local mucosa It is said that the mucosal immune system that exists in the region plays an important role in defense against infection.

  The mucosal immune system is known to have a unique control mechanism different from that of the systemic immune system, and is known to play a role in determining whether an antigen is favorable to the self or should be excluded. ing. In other words, in addition to the so-called “positive immune response” such as the elimination of foreign antigens and pathogenic microorganisms, the systemic immune system is made unresponsive to protein antigens orally ingested as food. It also controls the so-called “negative immune response”.

  For this reason, it is considered that by activating the mucosal immune system, in addition to the prevention and treatment of various diseases and infectious diseases, it is possible to sensitize allergies and to treat and prevent autoimmune diseases.

  As a food material having such a property that can activate the mucosal immune system, the following Patent Document 2 discloses a mucosal immune activation composition containing fructooligosaccharide as an active ingredient.

Patent Document 3 below discloses a mucosal immunity stimulating composition containing cyclofructan as an active ingredient, such as cyclic inulohexaose and cyclic inuloheptaose.
Japanese Patent Laid-Open No. 2001-322942 JP 2003-201239 A JP 2005-179195 A Edited by Hiroshi Seino, Hiromichi Ishikawa, Hiroshi Nakura, "Mucosal Immunity" pp.2-30 (2001)

  Lymphoid tissue on the local mucosal surface, called mucosa-associated lymphoid tissue, plays a central role in the mucosal immune system, and in particular, Peyer's plate, a typical mucosa-associated lymphoid tissue of the intestinal tract. patch) is a collection of all immunocompetent cells essential for induction and control of mucosal immunity, such as Th1-type / Th2-type CD4 + T cells, CD8 + T cells, IgA precursor B cells, dendritic cells, macrophages, etc. It is considered to have an important role as a control tower in controlling immunity. Therefore, if the formation of Peyer's patches can be enhanced, it is considered that the mucosal immune system can be more effectively activated.

  The indigestible saccharide disclosed in Patent Documents 2 and 3 activates the mucosal immune system by enhancing the production ability of secretory IgA, antibody secretory factor (pIgA), etc. Was unknown.

  Further, none of the above-mentioned Patent Documents 1 to 3 discloses a Peyer plate forming action.

  Accordingly, an object of the present invention is to provide a Peyer's patch formation inducing composition that can induce the formation of Peyer's patches and can be safely and easily ingested by the human body.

  As a result of intensive studies to achieve the above object, the present inventors have found that hemicellulose and / or a partially decomposed product thereof have an action of inducing the formation of Peyer's plate, and have completed the present invention. .

That is, Peyer's patches formed inducing composition of the present invention, the corn hulls from hemicellulose A partial hydrolyzate of hemicellulose obtained by enzymatic treatment, a weight average molecular weight of 30,000 to 80,000, arabinose And arabinoxylan containing xylose, galactose, glucose and glucuronic acid as constituent sugars .

As an active ingredient, a partial decomposition product of hemicellulose arabinoxylan Peyer's patches forming inducing composition of the present invention is highly safe, it can be easily taken orally. And because it can induce the formation of Peyer's patch, which plays a central role in the mucosa-related lymphoid tissue, it can activate the mucosal immune system more effectively, prevent and treat various diseases and infectious diseases, Effects such as desensitization and treatment / prevention of autoimmune diseases can be obtained.

Also, as an active ingredient of the Peyer's patches forming inducing composition of the present invention, a partial hydrolyzate of hemicellulose arabinoxylan is moderately degraded hemicellulose is under mild conditions, and low molecular weight without impairing the physiological activity effect Te, that has improved absorbent in the living body.

The active ingredient of the Peyer's board formation-inducing composition of the present invention, arabinoxylan, which is a partial decomposition product of hemicellulose, has a Peyer's board formation-inducing effect and is water-soluble and used as a raw material for pharmaceuticals, foods, drinks, feeds, feeds, etc. In addition, when added to these, the texture and flavor can be kept good.

Therefore, Peyer's patches formed inducing composition of the present invention is preferably used as a raw material for medicines.

Peyer's patches forming inducing composition of the present invention, highly safe, because it is an active ingredient arabinoxylan is a partial degradation product of the hemicellulose can be easily taken orally. And because it can induce the formation of Peyer's patch, which plays a central role in the mucosa-related lymphoid tissue, it can activate the mucosal immune system more effectively, prevent and treat various diseases and infectious diseases, Effects such as desensitization and treatment / prevention of autoimmune diseases can be obtained.

  Hemicellulose is a kind of dietary fiber. In particular, it is known that hemicellulose is contained in a large amount in cocoons and hulls of cereals such as rice, wheat, barley, and corn. Examples of the method for extracting hemicellulose from these hemicellulose-containing raw materials include, for example, extraction with alkali or acid, pressurization with an extruder or autoclave, extraction with hot water, extraction with an enzyme agent such as cellulase, or a combination thereof as appropriate. The method can be adopted.

In the present invention, Ru using corn hulls as hemicellulose-containing raw material. According to this, hemicellulose can be obtained with higher purity. Moreover, it is preferable to prepare hemicellulose by alkali extraction of the remainder from which starch, protein and, if necessary, lipids, minerals, etc. are removed from the hemicellulose-containing raw material, and starch, protein, and further if necessary It is particularly preferable to prepare a hemicellulose by alkali extraction of the remainder from which lipids, inorganic substances and the like have been removed.

  As a method for removing starch, protein, and, if necessary, lipids, minerals, and the like from the hemicellulose-containing raw material, any of enzyme treatment, chemical treatment, and physical treatment may be employed, or a combination thereof may be used as appropriate. Also good.

  As the enzyme treatment, a starch-degrading enzyme such as α-amylase or glucoamylase, a proteolytic enzyme such as protease, a lipolytic enzyme such as lipase, or a fibrinolytic enzyme such as cellulase is added to the hemicellulose-containing raw material, and the pH is 3 to 9, It is performed by operating under conditions of a temperature of 30 to 100 ° C.

  As chemical treatment, an aqueous solution of mineral acid or organic acid is added to the hemicellulose-containing raw material and heated under the conditions of pH 2 to 5, or a surfactant for food is added to the conditions of pH 3 to 8 It is carried out by heat treatment below.

  The physical treatment is performed by sieving the hemicellulose-containing raw material with a pulverizer such as a homogenizer or a hammer mill, and then sieving.

In the present invention, obtained by partial degradation by further enzymatic treatment hemicelluloses which are prepared as described above, shall be the active ingredient arabinoxylan is a partial degradation product of the hemicellulose.

Hemicellulose is, when the outer skin of corn as a raw material can be prepared, for example, as follows.

  Preferably, 80 to 95 parts by mass of water is added to 5 to 20 parts by mass of corn hulls that have been subjected to the above pretreatment (treatment for removing starch, protein, and if necessary, lipids, minerals, etc.), and an alkali compound is added. Add and adjust to pH 10-13, stir and mix at 80-140 ° C. for 0.5-10 hours to extract hemicellulose. There is no restriction | limiting in particular as an alkali compound, For example, calcium hydroxide and sodium hydroxide are mention | raise | lifted. The alkali compound is preferably added in the form of an aqueous solution in advance.

  The solution is then filtered to collect the filtrate and the extract is neutralized with acid. There is no restriction | limiting in particular as an acid, An organic acid and an inorganic acid can be used.

  This neutralized solution can be used as a crude product of hemicellulose by decolorizing and desalting, as it is, or by concentrating, or further pulverizing by spray drying or freeze drying.

  Furthermore, the protein precipitated by neutralization is separated and removed from the neutralized solution by means such as centrifugation, and the supernatant is dialyzed, ion exchange resin treated, ion exchange membrane treated, ultrafiltered as necessary. Hemicellulose of arbitrary purity can be obtained by processing alone or in combination as appropriate, such as membrane treatment, alcohol purification, and filter media treatment. In addition, when other hemicellulose-containing raw materials are used as raw materials, they can be prepared by the same method as described above.

  Moreover, the partial decomposition product of hemicellulose used in the present invention can be prepared, for example, as follows when corn hull is used as a raw material.

After clarifying filtration the resulting extract skin of corn and alkali extraction as described above, it intends rows reaction by hemicellulose degrading enzymes. By performing the treatment with hemicellulose-degrading enzyme, the molecular weight can be further reduced, the absorbability in the living body is high, and the bioactive effect is easily exhibited.

  As the hemicellulose-degrading enzyme used in the above reaction, commercially available enzyme preparations such as hemicellulase, xylanase, cellulase, and pectinase having hemicellulose-degrading activity can be used, and the alkali side described in Japanese Patent Publication No. 50-13357 is used. Preferred examples include alkaline xylanase “Celzyme” (trade name, manufactured by Nippon Shokuhin Kako Co., Ltd.) and “Pectinase GL” (trade name, manufactured by Amano Enzyme Co., Ltd.) having an optimum pH. Moreover, about the hemicellulose degradation conditions by the said hemicellulose decomposing enzyme, it can set suitably according to the characteristic of the enzyme formulation to be used.

  For example, when alkaline xylanase “cellzyme” is used as the hemicellulose-degrading enzyme, the extract obtained by alkali extraction of corn hulls as described above is clarified and filtered, and then adjusted to pH 50-60 It is preferable to react by adding xylanase at a temperature of ° C. The amount of xylanase added is preferably about 0.0001 to 10 units per gram of the solid content of the extract, and the reaction time is preferably about 3 to 96 hours. The titer of xylanase is measured by an enzyme that produces a reducing sugar corresponding to 1 μmol of xylose per minute under the reaction conditions of pH 7 and 60 ° C. using hemicellulose obtained by alkali extraction from corn hull as a substrate. The amount was taken as 1 unit.

  For example, when “pectinase GL” of pectinase is used as the hemicellulose-degrading enzyme, the amount of pectinase added is 0.01 to 1 g of the solid content of the extract under the conditions of pH 3 to 5 and 50 to 60 ° C. The reaction time is preferably about 3 to 96 hours.

  After the treatment with the enzyme is completed, the enzyme is deactivated by heating or the like, and then the reaction solution is decolorized, desalted, concentrated, or further dried to obtain a partially decomposed product of hemicellulose.

  Moreover, after preparing the solution containing 3-30 mass% of hemicellulose refine | purified as mentioned above, adjusting pH as needed, performing enzyme reaction like the above, and inactivating an enzyme, Solid-liquid separation may be performed by centrifugation or the like, and the supernatant liquid may be concentrated and dried. In addition, hemicellulose partial degradation products prepared by treating hemicellulose obtained from corn hulls with hemicellulose-degrading enzyme by the above-described method are, for example, “eclipse arabinoxylan” (trade name, Nippon Shokuhin Kako Co., Ltd.). Such as a commercial product can be used in the present invention. The “eclipse arabinoxylan” is a partially decomposed hemicellulose having a weight average molecular weight of about 50,000 to 60,000.

Which is the active ingredient of the present invention, a partial hydrolyzate of hemicellulose arabinoxylan, a weight average molecular weight of 3 0,000～80,000. The partially decomposed product of hemicellulose having such a weight average molecular weight has a low viscosity of 1 to 20 cps (B-type viscometer, 60 rpm, 25 ° C.) of a 5% by mass aqueous solution. When added to, the texture and flavor can be kept good. When the weight average molecular weight of the partially degraded hemicellulose exceeds 100,000,000, the viscosity becomes so high that it is difficult to add to raw materials for foods and drinks. This is not preferable because the active effect may be lost.

Then, as an active ingredient of the present invention, a partial hydrolyzate of hemicellulose arabinoxylan are those containing A arabinose, xylose, galactose, glucose and glucuronic acid as constituent sugars.

Peyer's patches forming inducing composition of the present invention, if example embodiment the aqueous solution containing the arabinoxylan is a partial degradation product of the hemicellulose can be commercialized as a concentrated solution or dry powder.

Among them, arabinoxylan is a partial degradation product of the hemicellulose derived from corn hulls, non-toxic, harmless nondigestible carbohydrates, has a slight sweetness, clear liquid tinged slightly yellowish or yellowish A white powder. And even when it is a dry powder, it dissolves easily in water, is excellent in heat resistance and acid resistance, is appropriately colored by heating, hardly harms other mixed materials, has low hygroscopicity and is in powder form However, since it has adhesion and anti-caking properties, it can also be used as a tablet in combination with a binder such as pullulan, hydroxyethyl starch, and polyvinylpyrrolidone. Furthermore, taste improvement, flavor improvement, osmotic pressure regulation, shaping, moisture retention, viscosity, anti-caking, aroma retention, discoloration prevention, stability, prevention of other sugar crystallization, starch aging prevention, protein These effects can also be expected because they have properties such as denaturation prevention and lipid degradation prevention.

Note that the arabinoxylan is a partial degradation product of the hemicellulose, starch that has not been removed, protein, lignin, cellulose, ash or the like may be contained.

  The Peyer's board formation-inducing composition of the present invention may be a saccharide (sucrose, starch syrup, flour koji, glucose, fructose, maltose, isomerized sugar, lactose, honey, coupling sugar, fructosyl oligosaccharide), if necessary. Sugar alcohols (erythritol, sorbitol, maltitol, xylitol, mannitol, lactitol, reduced xylooligosaccharide, reduced glucose syrup, etc.), high-intensity sweeteners (aspartame, alitame, saccharin, glycyrrhizin, stevioside, rebaudioside, sucralose, acesulfame K, etc. ), Sweeteners (glycine, alanine, etc.), enhancers (dextrin, starch, etc.), flavoring agents, coloring agents, flavoring agents, reinforcing agents, emulsifiers, antioxidants, UV inhibitors, medicinal ingredients, etc. You can also.

The effective intake of Peyer's patches forming inducing composition of the present invention, an adult per day, the arabinoxylan is a partial degradation product of the hemicellulose, it is preferable to 1~10g ingested in terms of solid content, is 3~5g More preferred.

Arabinoxylan, which is a partially decomposed product of hemicellulose, can be used as a raw material for pharmaceuticals, foods and drinks, feeds or feeds. Therefore, it is preferable that the Peyer's board formation induction composition of this invention is used as a raw material of a pharmaceutical.

  In the case of a pharmaceutical product, examples of the form include dosage forms such as liquids, powders, tablets, pills, fine granules, granules, capsules, jelly, chewable and paste.

  EXAMPLES Hereinafter, although this invention is demonstrated concretely based on an Example, this invention is not limited to these specific examples.

  In each of the following test examples and production examples, as the partially decomposed hemicellulose, “sun eclipse arabinoxylan” (trade name, manufactured by Nippon Shokuhin Kako Co., Ltd.) manufactured by the above-described method using corn hull as a raw material is used. It was. The molecular weight distribution of this hemicellulose partial decomposition product is shown in FIG. 1, and the analysis results of the constituent sugars are shown in Table 1, respectively.

(Test Example 1)
18 4-week-old female mice (C3H / Hej) were preliminarily raised for 7 days, and then divided into 3 groups (6 mice per group). The product dissolved in (1) was orally administered (50 mg / kg per dose) continuously for 21 days using a stomach tube. In test group 2, the hemicellulose partial decomposition product dissolved in distilled water was orally administered (150 mg / kg per dose) for 21 days using a stomach tube. Further, distilled water was administered to the control group in the same manner. In addition, food and water intake were free during the test period.
The general symptoms of the animals in each group during the study period were good, and there were no animals that died or were slaughtered.
After the 21-day test, the tested mice were laparotomized under Nembutal anesthesia. After necropsy, the pyloric region, Treitz's ligament, and the upper cecum were ligated and removed, and the number of Peyer's plates up to the duodenum, jejunum and ileum were measured. The measured value was expressed as an average value ± standard deviation, and the significance test was performed by student's t-test, and the presence or absence of a significant difference was determined at a risk rate of p <0.05 and p <0.01. The results are shown in Table 2.

  As can be seen from Table 2, in the test groups 1 and 2 to which the hemicellulose partial degradation product was administered, the number of Peyer's plates increased significantly (p <0.05) compared to the control group, and the numerical value was an increase in dose There was a tendency to increase in response.

At the time of dissection, the spleen was removed at the same time, and suspended cells were prepared by a conventional method. Adjust the number of cells with RPMI-1640 medium supplemented with 10% FCS, dispense 1 ml each into a 24-well plate at 2 × 10 ⁶ / ml / well, and then add ConA to a final concentration of 5 μg / cc / well. Then, the cells were cultured for 24 hours in a 37 ° C., 5% CO ₂ incubator. After the culture, the supernatant was collected, and IL-2, IL-4, IL-12, and IFN-γ were measured using a Biosource Cytoscreen kit and a plate reader system. Each measured value was expressed as an average value ± standard deviation, and the significance test was performed by student's t-test, and the presence or absence of a significant difference was determined at a risk rate of p <0.05 and p <0.01. The results are shown in Table 3.

  From Table 3, the test groups 1 and 2 to which the hemicellulose partial degradation product was administered suggested a dose-dependent increase-promoting action of cytokine (IL-2, IL-4 and IFN-γ) production ability against ConA stimulation of spleen cells. For IL-2 and IL-4, a significant increase (p <0.05) was observed with respect to the control group.

(Test Example 2)
7 pigs (L × W) with an average weight of 60 kg at 100 days of age were divided into two groups of 7 each, and the control group was given only the commercially available mixed feed, whereas the test group was partially decomposed into the commercially available mixed feed for the test group The animals were fed with 0.5% of the weight of the pig and fed for 60 days. From the feed amount, the remaining feed amount, and the weight gain during the 60-day test period, the average feed intake, daily weight gain, and required feed weight required for weight gain of both groups were calculated. The results are shown in Table 4.

  In the test group in which the hemicellulose partial degradation product was added to the commercially available mixed feed, it was confirmed that the feed intake, daily weight gain, and required feed amount were better fattening results than the control group. In addition, the health status of the pigs during the breeding period was good, and in particular, the fecal odor tended to be reduced in the test group. The intestinal immune system is activated by ingesting hemicellulose partial degradation products, and this is considered to be the effect of working on infection protection and the like.

(Production Example 1) [Biscuits]
All the ingredients in Table 5 were mixed in a mixer ball, mixed with a bread hook, and then placed in a refrigerator at 5 ° C., and the dough was spread to a thickness of 2 mm and formed into a 4 × 4 cm square. By adding the hemicellulose partial degradation product, no noticeable changes or problems were found except that the dough was slightly hardened. The dough was roasted at 165 ° C. for 15 minutes to obtain a bird biscuit. The hard biscuits obtained after firing had a yellowish brown color and a glossy appearance, and the inner layer became tight and tended to become hard. In terms of palatability, there were no particular problems with mouth feel, mouthfeel, sweetness, etc., and the flavor was fragrant and preferred.

(Production Example 2) [Jelly]
Table 6 shows the composition. The granulated sugar and the hemicellulose partial decomposition product were dispersed in half the amount of water, heated, boiled, and gelatin swollen with the remaining water was added and re-boiled. Immediately cooled, orange juice was added, and when it reached 50 ° C., it was poured into a mold and cooled and hardened in a refrigerator. The hemicellulose partial decomposition product has good dispersion solubility, and no problem in production was observed. Moreover, the transparency after dissolution was good, and no remarkable change or problem was found in the gel characteristics and gel strength of the tissue. In addition, there was no particular difference in palatability.

(Production Example 3) [Fruit drink]
In 1,000 ml of orange juice, 10 g of a partially degraded hemicellulose was added and dissolved. There was no particular problem with solubility and transparency, and it was a fruit juice drink with a rich taste.

(Production Example 4) [Tea drink]
10 g of hemicellulose partial degradation product was added to 1,000 ml of oolong tea and dissolved. There was no particular problem in solubility and transparency, and there was no sense of incongruity compared to ordinary tea drinks.

It is a graph which shows the molecular weight distribution of the hemicellulose partial decomposition product used in the Example of this invention.

Claims (2)

Corn hulls from hemicellulose A partial hydrolyzate of hemicellulose obtained by enzymatic treatment, a weight average molecular weight of 30,000 to 80,000, arabinose, xylose, galactose, as constituent sugars glucose and glucuronic acid A Peyer's board formation-inducing composition comprising the contained arabinoxylan as an active ingredient. Peyer's patches forming inducing composition according to claim 1 Symbol placement is used as a raw material of medicines.