JP2020178684A - Composition for improving intestinal flora diversity - Google Patents

Abstract

To provide a composition effective in improving of diversity of intestinal flora, and a composition effective in preventing or improving diarrhea, constipation, obesity or the like.SOLUTION: A composition for improving intestinal flora diversity containing 3 types or more of hardly digestible carbohydrate material selected from oligosaccharide (note that chitin oligosaccharide is excluded) and/or dietary fiber. It is particularly preferable that oligosaccharide be raffinose, galacto oligosaccharide, isomalto oligosaccharide, lacto oligosaccharide, lactulose, xylo oligosaccharide or fructo oligosaccharide. It is particularly preferable that dietary fiber be inulin, guar gum decomposed product, psyllium seed gum, hardly digestible dextrin, isomalto dextrin, cellulose, β-glucan, glucomannan or agarose. The composition for improving intestinal flora diversity can be a nutritive composition, a food composition, a pharmaceutical composition or a feed composition.SELECTED DRAWING: Figure 1

Description

The present invention relates to a composition for improving intestinal flora diversity. More specifically, the present invention relates to a composition containing three or more indigestible carbohydrate raw materials selected from oligosaccharides and / or dietary fiber to improve the diversity of the intestinal flora.

Bacteria such as a wide variety of bacteria and fungi are resident in the intestines of humans and mammals, and the collection of these bacteria is called the intestinal flora (intestinal flora).
Examples of intestinal bacteria include bifidobacteria, lactic acid bacteria, enterococci, eubacterium, etc., which are generally considered to be good bacteria, and Bacteroides, Clostridium, Escherichia coli, Staphylococcus aureus, which are generally considered to be bad bacteria. There are bacterium, Proteus, Pseudomonas aeruginosa, etc. In addition, fungi include Candida, Saccharomyces and the like. However, commonly used names such as good bacteria or beneficial bacteria, or bad bacteria or non-beneficial bacteria, or taxonomic ranks such as phylum, class, order, family, genus, species, and strain are not clearly attached. It should be noted that the name is a general name that ignores differences in the presence or absence of pathogenicity and metabolic characteristics depending on the bacterial species and strain, except when the bacterial species name and strain name are specified. ..
Recent studies have gradually shown that the intestinal flora is a complex ecosystem in which a large number of 1000 species of bacteria in humans perform various functions while maintaining a balanced population. It has become clear to. Not only bacteria that are beneficial to the host, but also bacteria that belong to a group of bacteria that are generally considered to be bad bacteria have some kind of ecological status in the intestinal flora and play a unique role. All of these form the intestinal flora. The healthy diversity of the gut flora itself allows the establishment and maintenance of ecosystem equilibrium, stabilizes the balance of populations, and overgrowth or activity of certain flora. It is considered to be a factor that suppresses the conversion. When this balance is lost and diversity is reduced (called disbiosis), pathogens grow and only specific biotransforms, including putrefactive products, accumulate in the intestine, resulting in the host human being. And an undesired intestinal environment for mammals. Causes of disbiosis include eating habits, age, environment, stress, and administration of drugs such as antibacterial drugs. Decreased diversity of the intestinal flora is known to cause various health disorders such as diarrhea, constipation, obesity, infectious diseases, immune disorders, and cancer, and cannot be underestimated.

Therefore, nutritional compositions and food compositions for maintaining and improving the diversity of the intestinal flora are required.
Currently, inulin is known as a substance for increasing beneficial intestinal bacteria. Further, Patent Document 1 discloses a composition containing any one of agar, agarose, and agarooligosaccharide and adjusting the intestinal flora. Patent Document 2 discloses a composition and a method containing chitin oligosaccharide, galactooligosaccharide, raffinose and lactulose to improve the balance of the intestinal flora. Patent Document 3 discloses an intestinal-regulating food containing konjac flour and oligosaccharides. Patent Document 4 discloses a composition containing an oligosaccharide and activating good intestinal bacteria. Further, Patent Document 5 discloses a cellooligosaccharide-containing composition. Further, Patent Document 6 discloses that oligosaccharides are added to viable intestinal bacteria in order to improve the intestinal flora.

JP 2017-163980 Patent 6163619 Japanese Patent Application Laid-Open No. 9-299059 International release 2012/141256 International release 2007/037249 JP-A-7-265064

However, none of the above prior arts are sufficient to improve the diversity of the intestinal flora.
For example, Patent Document 1 focuses only on specific taxa such as the phylum Bacteroidetes and the phylum Bacteroidetes in the intestinal flora, and has the effect of changing the abundance ratio to the normal type or the lean type. However, the composition of bacterial species in each taxon and the effect on taxa other than those shown were not taken into consideration. Further, Patent Document 2 is a technique relating to a composition containing four types of oligosaccharides including chitin oligosaccharides for the purpose of improving the balance of the intestinal flora, but pays attention only to an increase in bifidobacteria capable of assimilating oligosaccharides. However, it has not been able to exert its effect on the bacterial group that cannot assimilate these oligosaccharides, and if a large amount is ingested, there is a risk of causing symptoms such as diarrhea. Further, Patent Document 3 also aims to increase bifidobacteria only, and Patent Documents 4 and 5 also list only bifidobacteria and lactic acid bacteria as good bacteria or useful bacteria, and other bacterial groups and intestines. We did not pay attention to the diversity of the entire endobacterium flora.
Patent Document 6 provides a composition for improving the intestinal flora by orally supplementing viable bacteria. In addition, although there is a description that oligosaccharides are contained in this composition, the effect of oligosaccharides is defined as "selective growth of bacteria in the composition", and the effect on the entire intestinal flora that has existed before ingestion. Is not considered and there is no mention of the diversity of the intestinal flora.

Under such circumstances, the present invention provides a composition containing a safe material, which is superior to the conventional composition in improving the diversity of the intestinal flora, rather than a specific bacterium or bacterial group. That is the issue.
Another object of the present invention is to provide a composition effective for prevention or improvement of diarrhea, constipation, obesity and the like.

As a result of repeated studies for a composition containing a safe material capable of improving the diversity of the intestinal flora, the present inventor has found that three or more indigestible substances selected from oligosaccharides and / or dietary fiber We have found that by using a composition containing a carbohydrate raw material, the diversity of the intestinal flora can be improved and the above-mentioned problems can be solved, and the present invention has been completed.

That is, the present invention provides the following composition for improving intestinal flora diversity.
(1) A composition for improving intestinal flora diversity, which comprises three or more kinds of indigestible carbohydrate raw materials selected from oligosaccharides (excluding chitin oligosaccharides) and / or dietary fiber.
(2) The above-mentioned (1), wherein the oligosaccharide is raffinose, stachyose, galactooligosaccharide, isomaltooligosaccharide, milk fruit oligosaccharide, lactulose, xylooligosaccharide, agarooligosaccharide, mannooligosaccharide or fructooligosaccharide. Composition.
(3) The composition according to (1) or (2) above, wherein the oligosaccharide is raffinose, galactooligosaccharide, isomaltooligosaccharide, milk fruit oligosaccharide, lactulose, xylooligosaccharide or fructooligosaccharide.
(4) Dietary fiber is inulin, pectin, processed pectin, guar gum, guar gum decomposition product, psyllium seed gum, karaya gum, tragant gum, arabic gum, resistant starch, resistant dextrin, isomaltodextrin, polydextrose, cellulose , Hemicellulose, soybean polysaccharide, β-glucan, glucomannan, galactomannan, chondroitin sulfate, hyaluronic acid, levan, lignin, alginic acid and salts thereof, agarose or chitosan (1) to (3) above. The composition according to any one of.
(5) The dietary fiber is inulin, guar gum decomposition product, psyllium seed gum, indigestible dextrin, isomaltodextrin, cellulose, β-glucan, glucomannan or agarose, as described above (1) to (4). ).
(6) The composition according to any one of (1) to (5) above, wherein the composition ratio of each component of the indigestible carbohydrate raw material is 4 to 67% by mass, respectively.
(7) The composition according to any one of (1) to (6) above, wherein the daily intake is 30% by mass (dry mass) or less of the daily meal amount.
(8) The composition according to any one of (1) to (7) above, which has a preventive and ameliorating effect on constipation, diarrhea and obesity.
(9) The composition according to any one of (1) to (8) above, which is a nutritional composition, a food composition, a pharmaceutical composition or a feed composition.

According to the present invention, a composition which is excellent in improving the diversity of the intestinal flora and is useful for preventing or improving constipation, diarrhea and obesity can be obtained.

It is a photograph which shows the result of Test Example 2. It is a graph which shows the result of Test Example 3. It is a graph which shows the result of Test Example 4.

Hereinafter, the present invention will be described in detail.
The composition of the present invention can be used for maintaining or improving the diversity of the intestinal flora.
In the present invention, the "intestinal flora" means a collection of various bacteria such as bacteria and fungi that coexist in the intestines of humans and animals.
In the present invention, the high diversity of the intestinal flora means that there are many types of bacteria such as bacteria and fungi that compose the intestinal flora, and the balance between them is good. In addition, the composition of the present invention can improve the intestinal environment by improving the diversity of the intestinal flora.
The types of bacteria and fungi constituting the flora whose diversity is improved by using the composition for improving the diversity of the intestinal flora of the present invention are not limited to, for example, the phylum Bacteroidetes (Proteobacteria). Proteobacteria, Parabacteroides, Prebotera, Paraprebotera, Alloprebotera, Aristipes, etc.), Actinobacteria (Bifidobacterium, etc.), Fermicutes (Basilas, Elyciperotrics, Lacnospira , Crostridium, Eubacterium, Luminococcus, Phycaribacterium, etc.), Proteobacteria (Alpha Proteobacteria, Beta Proteobacteria, Gamma Proteobacteria, Delta Proteobacteria, etc.), Belcomicrovia Bacteria belonging to the phylum (Proteobacteria, etc.), Proteobacteria, etc. can be mentioned.

In addition, the composition of the present invention can be used for prevention, amelioration, and treatment of gastrointestinal symptoms. In this case, the gastrointestinal symptom includes, but is not limited to, constipation, diarrhea, abdominal pain, intestinal gas retention, and the like. Furthermore, the compositions of the present invention are also useful for the prevention, amelioration and treatment of obesity.
The composition of the present invention having such an effect can be used as a nutritional composition, a food composition, a pharmaceutical composition, a feed composition and the like.

The composition of the present invention is characterized by containing three or more kinds of indigestible carbohydrate raw materials selected from oligosaccharides (excluding chitin oligosaccharides) and / or dietary fiber. In the present invention, "oligosaccharide and / or dietary fiber" includes a case where it contains 3 or more kinds of oligosaccharides, a case where it contains 3 kinds or more of dietary fiber, and a case where it contains 3 or more kinds of oligosaccharide and dietary fiber in combination. Including.
Further, in the present invention, the "indigestible carbohydrate raw material" preferably contains 3 to 25 kinds of oligosaccharides and / or dietary fiber. When the number of types of oligosaccharide and / or dietary fiber is 2 or less, a sufficient effect of improving intestinal flora diversity cannot be obtained, and when the number of oligosaccharides and / or dietary fiber exceeds 25, the effect is not so different from 25 or less.

In the present invention, by increasing the diversity of the intestinal flora, it is possible to suppress the occurrence of harmful events such as loosening of the abdomen, and at the same time, the pH in the intestine is appropriately lowered to generate harmful substances such as ammonia. Can be suppressed and the intestinal environment can be improved.
In addition, by increasing the diversity of the intestinal flora according to the present invention, it is possible to increase the amount of short-chain fatty acids produced while suppressing the excessive production of lactic acid. Increased production of short-chain fatty acids provides nutrition to intestinal epithelial cells, promotes normal physiological activity of the intestinal tract, suppresses colon cancer, suppresses excessive immune response by regulatory T cell induction, and further Can obtain various desirable effects in health maintenance, disease prevention or disease management such as maintenance of energy metabolism homeostasis in liver, adipose tissue, muscle and the like.

The oligosaccharides contained in the composition of the present invention are not limited to these, but raffinose, stachiose, soybean-derived oligosaccharides, citrus-derived oligosaccharides, galactooligosaccharides, isomaltose, isomalttriose, panose, isomalt oligosaccharides, milk Fruit oligosaccharides, lactooligosaccharides, xylooligosaccharides, agarooligosaccharides, mannooligosaccharides, kestose, nistose, fructosylnistos, fructo-oligosaccharides and the like are preferable, and raffinose, galactooligosaccharides, isomaltoligosaccharides, milk fruit oligosaccharides, lactulose and xylooligosaccharides Alternatively, fructooligosaccharides are preferred. However, as an example of the oligosaccharide contained in the composition of the present invention, chitin oligosaccharide is not included.

The dietary fiber contained in the composition of the present invention is not limited to these, but is physically limited to levan, agricultural product-derived inulin, enzyme-synthesized inulin, inulin-type fractane, citrus peel-derived pectin, apple-derived pectin, and fruit-derived pectin. , Chemically or enzymatically modified pectin processed products, guar gum, guar gum decomposition products, psyllium seed gum, karaya gum, tragant gum, arabic gum, agricultural product-derived resistant starch, physical and chemical to ordinary starch Indigestible starch obtained by imparting indigestibility by a target or enzymatic chemical method, indigestible dextrin, isomalt dextrin, polydextrose, powdered cellulose, crystalline cellulose, hemicellulose, soybean-derived water-soluble polysaccharide, yeast-derived β-glucan, fungal-derived β-glucan, barley-derived β-glucan, agricultural product-derived β-glucan, agricultural product-derived glucomannan, agricultural product-derived galactomannan, cacao bean-derived lignin, plant-derived lignin such as rice family, alginic acid and its salts, Agarose, chitosan, chondroitin sulfate, hyaluronic acid and the like are preferable, and inulin, guar gum decomposition products, psyllium seed gum, indigestible dextrin, isomalto dextrin, cellulose, β-glucan, glucomannan or agarose are particularly preferable.

The composition ratio of each component of the indigestible carbohydrate raw material in the composition of the present invention is preferably 4 to 67% by mass of each component. When the composition ratio of each component is less than 4% by mass, the effect of a small amount of components is less likely to appear, and when it exceeds 67% by mass, the growth of only some bacteria is promoted, so that the intestinal flora as a whole In addition to the possibility of hindering the maintenance of diversity, the composition ratio of other components may be relatively reduced, and the effects of a small number of components may be less likely to appear as described above.

When the composition of the present invention is used as a nutritional composition, a food composition, or a feed composition, other nutritional components may be contained. Examples of other nutritional components include proteins, fats, carbohydrates, vitamins, minerals and the like. The type may be any of those usually contained in the nutritional composition, the food composition, and the feed composition.

Further, when the composition of the present invention is used as a pharmaceutical composition, it may contain other pharmaceutical ingredients. The other pharmaceutical component may be a component having an intestinal flora improving action or a component having other medicinal effects, and the type is not limited.

The composition of the present invention may contain components such as excipients, emulsifiers, stabilizers, pH adjusters, and fragrances. These types are not particularly limited, and those usually used in nutritional compositions, food compositions, pharmaceutical compositions, and feed compositions can be appropriately used.

The form of the composition of the present invention is not particularly limited, and any form may be used as long as it is in the form of a normal nutritional composition, food composition, pharmaceutical composition and feed composition. For example, it can be a powder, a liquid, a solid, a jelly, or the like.
The composition of the present invention can be produced by a conventional production method for each of a nutritional composition, a food composition, a pharmaceutical composition, and a feed composition. The administration target in the case of a feed composition is not particularly limited, and examples thereof include pets, livestock, poultry, and fish farming.

The route of administration of the composition of the present invention is not particularly limited, and may be oral administration or tube administration using a nasal, gastric fistula, intestinal fistula tube or the like. The number of administrations can be divided into once to several times a day and given as appropriate.

The intake or dose of the composition of the present invention is 1 to 30% by mass (dry mass), preferably 1 to 20% by mass (dry mass), and more preferably 1 to 10 of the daily dietary amount (dry mass). It is preferably mass% (dry mass). In addition, the intake amount or the dose can be appropriately determined according to the physical condition, pathological condition, body weight, age and the like. Even if it is administered in excess of 30% by mass (dry mass) of the daily meal amount, no further effect can be expected to improve the flora diversity. In addition, administration of an excessive amount may excessively promote a feeling of fullness, which may hinder the intake of an appropriate amount of other nutrients such as proteins and lipids.
The amount of each component contained in the composition of the present invention is preferably less than or equal to the acceptable daily intake (ADI).

Hereinafter, the present invention will be described in more detail with reference to Examples and Test Examples. Unless otherwise specified, "%" in Examples and Test Examples means "% by mass".
(Example 1)
Reeves PG et al. J Nutr. 1993, 123: 1939-1951. AIN-93 purified diets for laboratory rodents: Final report of the American Institute of Nutrition ad hoc writing committee on According to the reformulation of the AIN-76A rodent diet.), The compositions of "Example 1", "Comparative Example 1" and "Comparative Example 2" having the compositions shown in Table 1 below were prepared.
As shown in Table 1, the composition of "Example 1" contains 25 kinds of dietary fiber components and oligosaccharide components in a total amount of 6.8% by mass. "Comparative Example 2" contains 6.8% by mass of inulin, which has been conventionally used for improving the intestinal environment, as a dietary fiber component, as in Example 1. "Comparative Example 1" had the same composition as AIN-93G and contained 5% by mass of cellulose as dietary fiber, and was used as a control.
Of the dietary fiber components and oligosaccharide components contained in "Example 1" and "Comparative Example 2", the amount blended in excess of 5% by mass, which is equivalent to cellulose in the AIN-93G composition, is one of cornstarch. It was prepared by replacing the part. The same applies to the following examples and test examples.

(Test Example 1)
・ Improvement of intestinal flora diversity (intestinal flora diversity index)
Male C57BL / 6J mice were acclimatized from 6 weeks of age for 1 week, and the compositions of "Example 1", "Comparative Example 1" and "Comparative Example 2" were fed as feed for 4 weeks. The number of animals constituting each group was 4 or 5 (the same applies to the following test examples unless otherwise specified).
After ingesting each composition for 4 weeks, mouse feces were collected to extract DNA, and the V3 to V4 regions of the 16S rRNA gene of enterobacteria were amplified by the PCR method, and the nucleotide sequence of the amplified product DNA was obtained as follows. The analysis was performed using a generation sequencer Miseq (manufactured by Illumina Co., Ltd.). An OTU (Operational Taxonomy Unit) was created by clustering similar nucleotide sequences, the sequence of each OTU was collated with a database, the most recently related bacterial species were assigned, and the number of detected bacterial species was determined.
In addition, the abundance ratio of the corresponding bacterial species was calculated from the abundance ratio of OTU, and the diversity index of Shannon-Wiener was calculated by applying the following formula (Dean-Ross et al. Appl Environ Microbiol. 1989). , 55 (8): 2002-2009. Bacterial community structure and function along a heavy metal gradient.).

Table 2 shows the number of detected bacterial species and the average value of the obtained Shannon-Wiener diversity index.

As is clear from the results shown in Table 2, the mice fed with the composition of "Example 1" were compared with the mice fed with the compositions of "Comparative Example 1" and "Comparative Example 2". The number of bacterial species and the diversity index were very high.
From this result, it was found that the composition of "Example 1" significantly increases the diversity of the intestinal flora of mice by containing 25 kinds of dietary fiber and oligosaccharides.

(Example 2)
Similar to the case of Example 1, the compositions of "Example 2", "Comparative Example 1" and "Comparative Example 3" having the compositions shown in Table 3 were prepared according to the AIN-93G composition.
The composition of "Example 2" contains the same 25 kinds of dietary fiber and oligosaccharides as in "Example 1" in a total mass ratio of 10% by mass as shown in Table 3. The composition of "Comparative Example 1" is the same as that used in Test Example 1. "Comparative Example 3" contains 10% by mass of inulin as a dietary fiber component.

(Test Example 2)
-Normalization of fecal properties Male Sprague-Dawley rats were acclimatized from 6 weeks of age to 1 week, and the compositions of "Example 2" and "Comparative Example 3" were fed as feed for 2 weeks, and then each animal. A photograph of the stool was taken (Fig. 1).
As is clear from the results shown in FIG. 1, the stools of the rats fed with the composition of "Example 2" were normal stools, whereas the stools contained only inulin as dietary fiber "Comparative Example 3". The stools of rats fed with the composition of "" were loose stools or muddy stools. From this, it was found that the stool properties of rats can be normalized by using the composition of "Example 2".

(Test Example 3)
-Adjustment of intestinal pH Male Sprague-Dawley rats are acclimatized from 6 weeks of age to 1 week, and the compositions of "Example 2", "Comparative Example 1" and "Comparative Example 3" are ingested as feed for 2 weeks. After that, the contents of the cecum of each animal were collected, and the pH was measured using a compact pH meter B-212 (manufactured by HORIBA, Ltd.). The results are shown in FIG.
It is not desirable if the intestinal pH is too low or too high, but as is clear from the results shown in FIG. 2, the composition of "Example 2" containing 25 kinds of dietary fiber and oligosaccharide is dietary fiber. While containing the same concentration (10% by mass) of the indigestible carbohydrate raw material as the composition of "Comparative Example 3" containing one kind, the intestinal pH of the animal is moderately higher than that of the composition of "Comparative Example 3". Was reduced to. From this, it was found that the composition of "Example 2" was excellent in adjusting the intestinal pH.

(Test Example 4)
・ Effects on organic acid production in the intestinal tract (amount of lactic acid, butyric acid, propionic acid)
In this test example, the effects of the compositions of "Example 2", "Comparative Example 1" and "Comparative Example 3" on the production of lactic acid, butyric acid and propionic acid in the cecum of rats were confirmed.
Male Sprague-Dawley rats were acclimatized from 6 weeks of age to 1 week, and the compositions of "Example 2", "Comparative Example 1" and "Comparative Example 3" were fed as feed for 2 weeks.
After ingesting each composition, the contents of the cecum were collected, diluted and extracted with purified water, and then perchloric acid was added for deproteinization. Further, the filtered extract was subjected to ion elimination high performance liquid chromatography, the concentrations of lactic acid and short-chain fatty acids butyric acid and propionic acid were measured, and the amount of each organic acid per cecum was calculated by multiplying the weight of the contents of the cecum. did. The results are shown in FIG.
As is clear from the results shown in FIG. 3, the composition of "Example 2" contains the same concentration (10% by weight) of the indigestible carbohydrate raw material as the composition of "Comparative Example 3", while containing the rat cecum. It suppressed the excessive accumulation of lactic acid in the cecum and significantly increased the amount of short-chain fatty acids in the contents of the cecum.
From this result, when the composition of "Example 2" containing 25 kinds of dietary fiber and oligosaccharide was ingested, the composition of "Comparative Example 3" containing only one kind of dietary fiber even though the content was the same. It can be seen that the flora did not accumulate lactic acid and produced a large amount of short-chain fatty acids, as compared with the case of ingesting.
From this, it was found that the composition containing 25 kinds of dietary fiber and oligosaccharide had a higher ability to produce short-chain fatty acids than the composition containing 1 kind of dietary fiber.

(Examples 3 and 4)
The compositions of "Example 3", "Example 4" and "Comparative Example 3" having the compositions shown in Table 4 below were prepared.
As shown in Table 4, the composition of "Example 3" contains three kinds of dietary fiber components, inulin, guar gum decomposition product, and glucomannan, and "Example 4" contains dietary fiber components. It contains a total of four types, three types of inulin, guar gum decomposition product, and glucomannan, and one type of galactooligosaccharide as an oligosaccharide component. "Comparative Example 3" is the same as that shown in Table 3.

(Test Example 5)
・ Improvement of intestinal flora diversity by ingesting 3 or 4 mixed compositions (indicator of flora diversity)
After ingesting the compositions of "Example 3", "Example 4" and "Comparative Example 3" into rats in the same manner as in Test Example 1, the diversity of the intestinal flora was evaluated.
Table 5 shows the number of detected bacterial species and the average value of the obtained Shannon-Wiener diversity index.

As is clear from the results shown in Table 5, rats fed with the compositions of "Example 3" containing 3 types of dietary fiber and "Example 4" containing 4 types of dietary fiber and oligosaccharides Compared with the rats fed with the composition of "Comparative Example 3", the number of bacterial species was large and the diversity index was also high.
From this result, it was found that the compositions of "Example 3" and "Example 4" significantly increased the number and diversity of the intestinal flora of rats. Further, comparing the compositions of "Example 3" and "Example 4", "Example 3" containing only 3 types of dietary fiber contains 4 types of dietary fiber and oligosaccharides, as compared with the composition of "Example 3". It was found that the composition of "Example 4" has a higher effect of improving the diversity of the intestinal flora.

(Example 5)
The composition of "Example 5" having the composition shown in Table 6 below was prepared. The compositions of "Example 2" and "Comparative Example 3" displayed together with the composition of "Example 5" in Table 6 are the same as those shown in Table 3. The composition of "Example 2" contains 25 kinds of dietary fiber and oligosaccharide, and the composition of "Example 5" contains cellulose, inulin, guar gum decomposition product, psyllium seed gum, and isomaltodextrin as dietary fiber. , Β-Glucan, Glucomannan and Agarose.

(Test Example 6)
・ Improvement of intestinal flora diversity by ingestion of 8 and 25 mixed compositions (indicator of flora diversity)
In this test example, the compositions of "Example 2", "Example 5" and "Comparative Example 3" were ingested by rats using the same method as in Test Example 1, and then the intestinal flora. We evaluated the diversity of.
The average value of the obtained Shannon-Wiener diversity index is shown in Table 7.

As is clear from the results shown in Table 7, the composition of "Example 5" containing 8 kinds of dietary fiber is the same as the composition of "Example 2" containing 25 kinds of dietary fiber and oligosaccharide. Increased the diversity of the rat intestinal flora.
Comparing the compositions of "Example 2" and "Example 5", the composition of "Example 5" is a compounding type of dietary fiber and / or oligosaccharide as compared with the composition of "Example 2". Although the number was as small as about one-third, the diversity of the intestinal flora of rats was increased to the same value as in Example 2.
From this, it was found that the number of types of dietary fiber and / or oligosaccharide blended was about 3 to 25, and the same effect was obtained.

(Examples 6 to 10)
The compositions of "Example 6", "Example 7", "Example 8", "Example 9" and "Example 10" having the compositions shown in Table 8 below were prepared. The compositions of "Example 6" to "Example 10" contain seven kinds of dietary fiber components, cellulose, inulin, guar gum decomposition product, psyllium seed gum, indigestible dextrin, glucomannan and agarose, and an oligosaccharide component. As a whole, it contains a total of 8 types including 1 type of galactooligosaccharide. The blending amount of dietary fiber and oligosaccharide in each composition was 1% in "Example 6", 5% in "Example 7", 10% in "Example 8", and 20% in "Example 9". And "Example 10" was set to 30%.

(Test Example 7)
・ Improvement of intestinal flora diversity (intestinal flora diversity index)
In this test example, the diversity of the intestinal flora was evaluated after ingesting each composition in rats using the same method as in test example 1.
In order to examine the appropriate blending ratio of dietary fiber and oligosaccharide, each composition of "Example 6" to "Example 10" (blending amount is 1 to 30% by mass) is used to compose a mixture of 8 kinds. We confirmed the effective dose of the effect of improving intestinal flora diversity in rats by ingestion (index of flora diversity).
The average value of the obtained Shannon-Wiener diversity index is shown in Table 9.

As is clear from the results shown in Table 9 below, "Example 6" (1% combination), "Example 7" (5% combination), "Example 8" (10% combination), "Example 9". (20% formulation) and "Example 10" (30% formulation) were both able to increase the diversity of the rat intestinal flora.
Among these, each composition of "Example 6" (1% combination), "Example 7" (5% combination) and "Example 8" (10% combination) increased the diversity of the rat intestinal flora. It was particularly effective against.
From this, it was found that the composition of the present invention was effective at a blending amount of 1 to 30% by mass with respect to the amount of food (dry weight), and was particularly effective at 1 to 10% by mass.

(Test Example 8)
・ Prevention and improvement effect of obesity (index of intestinal flora associated with obesity)
Recent studies have shown that the relative abundance of bacteria belonging to the phylum Bacteroidetes is high, the relative abundance of bacteria belonging to the phylum Bacteroidetes is low, and these ratios (phylum Bacteroidetes / Bacteroidetes) are high. , It has been pointed out that it may cause weight gain and obesity (Turnbaugh PJ et al. Nature. 2006, 444: 1027-1031. An obesity-associated gut microbiome with increased capacity for energy harvest.).
Therefore, in this test example, for each composition of "Example 2", "Example 5" and "Comparative Example 3" shown in Table 6, each bacterial species in the rat intestinal flora obtained in Test Example 6 The above-mentioned index was evaluated using the relative abundance ratio data of, and the effect of the composition of the present invention on the prevention and amelioration of obesity was investigated. The average value of the obtained values is shown in Table 10.

As is clear from the results shown in Table 10, which is the composition of "Example 5" containing 8 kinds of dietary fiber and the composition of "Example 2" containing 25 kinds of dietary fiber and oligosaccharides? Also greatly increased the relative abundance ratio of Bacteroidetes and significantly reduced the relative abundance ratio of Famicutes / Bacteroidetes.
From this, it can be seen that the composition of the present invention is effective in preventing and ameliorating obesity.

(Example 11)
-Nutrition composition As the amount per 100 mL of the composition, 0.340 g of psyllium seed gum powder, 0.310 g of konjak glucomannan powder, 0.410 g of cellulose powder, 0.440 g of barley β-glucan powder, 0.440 g of guar gum decomposition product powder, Inulin powder 0.410 g, refractory dextrin 0.450 g, isomaltodextrin powder 0.450 g, milk casein 3.400 g, isolated soybean protein 1.666 g, tricapriline 0.750 g, soybean oil 0.699 g, perilla oil 0.180 g, palm oil 0.334 g, maltodextrin 14.858 g, purified sucrose 1.300 g, various vitamins, various minerals and various additives are dissolved in water for production and mixed well with stirring so as to be uniform. A pouch made of an aluminum laminate film was filled with 200 mL each and subjected to retort heat treatment to obtain 200 mL of a liquid nutritional composition. Moreover, 0.450 g of agar was used instead of indigestible dextrin to obtain a semi-solid nutritional composition.
By ingesting the obtained nutritional composition, a total of 3.25 g of dietary fiber and oligosaccharides can be ingested per 100 mL, and at the same time, proteins, lipids, carbohydrates, various vitamins and various minerals can be supplemented. it can.

(Example 12)
-Food composition 93% by weight of water for production was heated, agar powder 0.37% by weight, psyllium seed gum powder 0.34% by weight, konjak glucomannan powder 0.31% by weight, cellulose powder 0.41% by weight, 0.44% by weight of barley β-glucan powder, 0.44% by weight of guar gum decomposition product powder, 0.41% by weight of inulin powder and 0.45% by weight of isomaltodextrin powder were doubled into 2.0% of purified sucrose. It was added and dissolved, 0.50% by weight of citric acid and 0.37% by weight of sodium citrate were added for pH adjustment, and the mixture was stirred and mixed. After appropriately adding mixed water to make the total amount 100% by weight, 180 g per pouch is filled in an aluminum pouch with a spout, and after heat treatment in hot water, it is cooled and solidified to obtain 180 g of a jelly-like food composition. It was.
The obtained food composition contained a total of 8 types of dietary fiber and oligosaccharide in an amount of 3.17% by weight, and exhibited a good flavor as a jelly-like food.

(Example 13)
-Pharmaceutical composition Agar powder 7.4% by weight, psyllium seed gum powder 7.0% by weight, konjak glucomannan powder 7.6% by weight, cellulose powder 7.9% by weight, barley β-glucan powder 8.0% by weight , 7.9% by weight of guar gum decomposition product powder, 8.0% by weight of inulin powder and 7.9% by weight of isomaltodextrin powder, and 38.3% by weight of maltodextrin as a diluting agent were added to make it uniform in a powder mixer. The mixture was sufficiently mixed so that 8 g of each was filled with a three-way seal with an aluminum laminate film to obtain 8 g of a powdery composition.
The obtained composition contains 61.7% by weight of a total of 8 types of dietary fiber and oligosaccharide, and can be used as a pharmaceutical composition. Furthermore, various dosage forms of pharmaceutical compositions can be obtained by using other base components. In addition, these can also be used as a food composition or a nutritional composition.

Claims (9)

A composition for improving intestinal flora diversity, which comprises three or more kinds of indigestible carbohydrate raw materials selected from oligosaccharides (excluding chitin oligosaccharides) and / or dietary fiber. The composition according to claim 1, wherein the oligosaccharide is raffinose, stachyose, galactooligosaccharide, isomaltooligosaccharide, milk fruit oligosaccharide, lactulose, xylooligosaccharide, agarooligosaccharide, mannooligosaccharide or fructooligosaccharide. The composition according to claim 1 or 2, wherein the oligosaccharide is raffinose, galactooligosaccharide, isomaltooligosaccharide, milk fruit oligosaccharide, lactulose, xylooligosaccharide or fructooligosaccharide. Dietary fiber is inulin, pectin, pectin processed product, guar gum, guar gum decomposition product, psyllium seed gum, karaya gum, tragant gum, arabic gum, resistant starch, resistant dextrin, isomalt dextrin, polydextrose, cellulose, hemicellulose, The invention according to any one of claims 1 to 3, which is a soybean polysaccharide, β-glucan, glucomannan, galactomannan, chondroitin sulfate, hyaluronic acid, levan, lignin, alginic acid and a salt thereof, agarose or chitosan. Composition. The invention according to any one of claims 1 to 4, wherein the dietary fiber is inulin, guar gum decomposition product, psyllium seed gum, indigestible dextrin, isomalto dextrin, cellulose, β-glucan, glucomannan or agarose. Composition. The composition according to any one of claims 1 to 5, wherein the composition ratio of each component of the indigestible carbohydrate raw material is 4 to 67% by mass, respectively. The composition according to any one of claims 1 to 6, wherein the daily intake is 30% by mass (dry mass) or less of the daily meal amount. The composition according to any one of claims 1 to 7, which has a preventive and ameliorating effect on constipation, diarrhea and obesity. The composition according to any one of claims 1 to 8, which is a nutritional composition, a food composition, a pharmaceutical composition or a feed composition.

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