Butyric acid bacterium proliferation promoting agent, and food and pharmaceutical including the same

Abstract

To provide a synergistic or counteracting butyric acid bacterium proliferation-promoting agent which includes a combination capable of exhibiting an effect of further synergistically promoting the proliferation of butyric acid bacteria, compared to the effect observed when a nutritional component is ingested alone, or a combination capable of exhibiting an effect of promoting the proliferation of butyric acid bacteria by counteracting the inhibitory effect on proliferation observed when the nutritional component is ingested alone, by combining 1-kestose with the nutritional component.SOLUTION: A butyric acid bacterium proliferation-promoting agent according to the present invention is a synergistic or counteracting butyric acid bacterium proliferation-promoting agent including 1-kestose and a nutritional component. The nutritional component is one or more components selected from predetermined polyphenols, predetermined vitamins, predetermined lipids, predetermined amino acids, predetermined proteins, and predetermined minerals.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present invention relates to a butyric acid bacteria growth promoter, and to a food and a pharmaceutical product containing the same. [Background technology]

[0002] Butyric acid (butanoic acid, n-butanoic acid) is a straight-chain carboxylic acid with the molecular formula C4H8O2 and the rational formula CH3(CH2)2COOH. Butyric acid is one of the short-chain fatty acids that are major metabolic products of intestinal bacteria, and is known to keep the intestine weakly acidic, inhibit the growth of harmful bacteria (so-called bad bacteria), and provide various beneficial physiological functions in the host body. For example, butyric acid is known to be involved in the differentiation induction of regulatory T cells (Treg) that are responsible for immune tolerance in peripheral tissues, and in the promotion of the production of IL-10, a cytokine that suppresses inflammation and autoimmune responses (Non-Patent Document 1: Nature, 2013; 504(7480): 446-450, Non-Patent Document 2: Nutrients, 2011; 3(10): 858-876). Butyric acid is also known to be involved in regulating the production or secretion of hormones such as leptin and peptide YY (PYY), which have appetite suppressing effects, and glucagon-like peptide-1 (GLP-1), which acts as an incretin and has an effective effect on type 2 diabetes (Non-Patent Document 3: Journal of Veterinary Science, 2011;12(4):319-323, Non-Patent Document 4: Scientific Reports, 2018;8:74).

[0003] However, even a small amount of butyric acid gives off an unpleasant odor, so it is not practical to actively ingest it directly as a food ingredient. Therefore, it is desirable to produce butyric acid as a metabolic product by intestinal bacteria. Butyric acid is produced by butyric acid bacteria, which are known as useful bacteria (so-called good bacteria) in the intestinal flora. Butyric acid bacteria is a general term for bacteria that produce butyric acid, i.e., have the ability to produce butyric acid, and are also called butyric acid-producing bacteria. Among butyric acid bacteria, for example, Faecalibacterium duncaniae (formerly known as Faecalibacterium prausnitizii) belonging to the Clostridium leptum subgroup (Clostridium Cluster IV), Agathobacter rectalis (formerly known as Eubacterium rectale), Eubacterium bacteria, and Rosebria bacteria belonging to the Clostridium coccoides group (Clostridium cluster XIVa), can be exemplified as dominant bacteria in the human intestinal flora (Non-Patent Document 5: FEMS Microbiology Letters, 2009;294(1):1-8).

[0004] A practical and effective way to increase butyric acid in the intestines is to ingest butyric acid bacteria (probiotics) that can be consumed in food form, such as Clostridium butyricum, also known as Miyairi bacteria.

[0005] There is also a method of ingesting ingredients (so-called prebiotics) that reach the lower digestive tract without being decomposed or absorbed in the upper digestive tract and serve as a selective substrate for beneficial bacteria including butyric acid bacteria, promoting their growth or activating their metabolism. 1-Kestose is known as one of the prebiotics that has the effect of promoting the growth of butyric acid bacteria (Non-Patent Document 6: Gut Microbes, 2021; 13(1):1869503-1869503-20). [Prior art documents] [Non-patent literature]

[0006] [Non-licensed document 1] Y.Furusawa et al., Springer Nature (GB), Nature, 2013;504(7480):446-450 [Non-licensed document 2] Vinolo MA et al., MDPI (Multidisciplinary Digital Publishing Institute) (CH), Nutrients, 2011;3(10):858-876 [Non-licensed document 3] Soliman MM et al, Korean Society of Veterinary Science (KR), Journal of Veterinary Science, 2011;12(4):319-323

Non-licensed Document 4

Non-licensed Document 5

Non-licensed Document 6

Non-licensed Document 7

Non-licensed Document 8

[0007] Incidentally, foods containing specific nutritional components, which are collectively called health foods or called dietary supplements, nutritional supplements, supplements, etc., and which are consumed in the hope of replenishing the nutritional components or achieving specific functions, are now in widespread use. Examples of nutritional components contained in these foods include carbohydrates, dietary fiber, polyphenols, vitamins, lipids, amino acids, proteins, minerals, etc.

[0008] Among these nutritional components, for example, Non-Patent Document 7 (Food & Function, 2021;12(6): 8850-8866) describes that polyphenols contained in fruits may promote the growth of butyric acid bacteria. However, it has not yet been clarified whether each nutritional component has the function of promoting the growth of butyric acid bacteria. On the other hand, some nutritional components contain components that can suppress the initial growth of bifidobacteria, which are known as useful bacteria like butyric acid bacteria (Non-Patent Document 8: Acta Biochimica Polanica, 2015;62(4):895-901), and it is possible that some nutritional components contain components that can suppress the growth of butyric acid bacteria. [Means for solving the problem]

[0009] In light of the above background, the present inventors have investigated the effect of each nutritional component on the proliferation of butyric acid bacteria. Furthermore, they have also investigated the effect on the proliferation of butyric acid bacteria when these nutritional components are combined with 1-kestose, which is known to promote the proliferation of butyric acid bacteria. For example, in the case of medicinal components, even if the usefulness of each component is recognized individually, the effects may be antagonized and canceled out when used in combination. In the technical field to which the present invention pertains, even if the proliferation-promoting effect of butyric acid bacteria is recognized individually, it cannot necessarily be concluded that the combination of these components is useful.

[0010] Under these circumstances, as a result of investigation, it was found that there exist components whose growth-promoting effect on butyric acid bacteria confirmed when used alone is further manifested as a synergistic growth-promoting effect when used in combination with 1-kestose (details will be described later, but this effect will be defined as a synergistic growth-promoting effect), or whose growth-inhibiting effect on butyric acid bacteria confirmed when used alone is counteracted when used in combination with 1-kestose, resulting in a further growth-promoting effect on butyric acid bacteria (details will be described later, but this effect will be defined as a countervailing growth-promoting effect), which led to the present invention.

[0011] In other words, the present invention aims to provide a synergistic or countervailing growth promoter for butyric acid bacteria, including a combination of 1-kestose and a nutritional component, which can exert a synergistic growth-promoting effect on butyric acid bacteria when the nutritional component is ingested alone, or a combination in which the growth-inhibiting effect on butyric acid bacteria when the nutritional component is ingested alone is offset, resulting in a further growth-promoting effect on butyric acid bacteria, as well as foods and pharmaceuticals containing the same.

[0012] The present invention solves the above problems by the solution means described below as one embodiment.

[0013] One embodiment of the growth promoter for butyric acid bacteria of the present invention is a synergistic or countervailing growth promoter for butyric acid bacteria comprising 1-kestose and a nutritional component, characterized in that the nutritional component is one or more polyphenols selected from rutin, quercetin, catechin, hesperidin, daidzein, genistein, p-coumaric acid, caffeic acid, and vanillin.

[0014] Furthermore, one embodiment of the growth promoter for butyric acid bacteria according to the present invention is a synergistic or countervailing growth promoter for butyric acid bacteria comprising 1-kestose and a nutritional component, characterized in that the nutritional component is one or more vitamins selected from biotin, niacin, pantothenic acid, riboflavin, vitamin E, and vitamin D, derivatives of these vitamins, salts of the vitamins, and salts of the derivatives.

[0015] Furthermore, one embodiment of the butyric acid bacteria growth promoter of the present invention is a synergistic or countervailing growth promoter for butyric acid bacteria comprising 1-kestose and a nutritional component, characterized in that the nutritional component is one or more lipids selected from arachidonic acid, α-linolenic acid, and docosahexaenoic acid, as well as compounds having these fatty acids as constituent fatty acids.

[0016] Furthermore, one embodiment of the butyric acid bacteria growth promoter of the present invention is a synergistic or countervailing growth promoter for butyric acid bacteria comprising 1-kestose and a nutritional component, characterized in that the nutritional component is one or more amino acids or proteins selected from γ-aminobutyric acid and whey protein.

[0017] Furthermore, one embodiment of the butyric acid bacteria growth promoter of the present invention is a synergistic or countervailing growth promoter for butyric acid bacteria comprising 1-kestose and a nutritional component, characterized in that the nutritional component is one or more minerals selected from potassium, magnesium, and iron.

[0018] Furthermore, the growth promoter for butyric acid bacteria according to the present invention can also be a synbiotic containing butyric acid bacteria in addition to the 1-kestose and the nutritional components.

[0019] The food product according to the present invention is provided as a food product containing the butyric acid bacteria growth promoter according to the present invention.The pharmaceutical product according to the present invention is provided as a pharmaceutical product containing the butyric acid bacteria growth promoter according to the present invention. Effect of the Invention

[0020] According to the present invention, the combined use of 1-kestose and a specified nutritional component can provide a synergistic or countervailing effect on the growth of butyric acid bacteria. [Brief description of the drawings]

[0021] [Figure 1] 1A, 1B, and 1C are explanatory diagrams illustrating the synergistic growth-promoting effect of butyric acid bacteria and the countervailing growth-promoting effect of butyric acid bacteria according to the present invention. FIG. 1A is a graph showing an example of the growth amount of butyric acid bacteria. FIG. 1B is a graph showing an example of the synergistic growth-promoting effect of butyric acid bacteria. FIG. 1C is a graph showing an example of the countervailing growth-promoting effect of butyric acid bacteria. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0022] Hereinafter, an embodiment of the present invention will be described.

[0023] The butyric acid bacteria growth promoter according to the present embodiment is a synergistic or countervailing butyric acid bacteria growth promoter containing 1-kestose and a specific nutritional component. 1-kestose may be abbreviated as "Kes". First, the synergistic and countervailing butyric acid bacteria growth promotion effects according to the present invention are defined. FIG. 1A shows an example of the amount of growth of butyric acid bacteria at a certain time point (e.g., 24 hours, 48 ​​hours, etc.) after the start of culture when butyric acid bacteria are cultured in a specific medium, as a turbidity increase amount representing the net turbidity obtained by subtracting the turbidity measurement value at the start of culture from the turbidity measurement value of the medium at that time point.

[0024] Point a represents an example of culture using only basal medium components. Point b represents an example of culture using basal medium components with the addition of a specified nutrient component. Point b shows a larger increase in turbidity than point a, and the nutrient component has a growth-promoting effect on butyric acid bacteria. Point c represents an example of culture using basal medium components with the addition of 1-kestose. Point c shows a larger increase in turbidity than point a, and 1-kestose has a known growth-promoting effect on butyric acid bacteria. Point d represents an example of culture using basal medium components with the addition of 1-kestose and the specified nutrient component.

[0025] In this case, the difference in the turbidity increase between points a and b, represented by the line a and b, represents the growth-promoting effect of the nutrient component alone on butyric acid bacteria, in other words, the growth-promoting effect of the nutrient component in the absence of 1-kestose. In contrast, the difference in the turbidity increase between points c and d, represented by the line cd, represents the growth-promoting effect of the combined use of 1-kestose and a nutrient component, in other words, the growth-promoting effect of the nutrient component in the presence of 1-kestose. Therefore, by comparing the effect represented by the line a and the effect represented by the line cd, it is possible to evaluate whether or not the combined use of 1-kestose and a nutrient component can further exert a synergistic growth-promoting effect on butyric acid bacteria by the combined use of 1-kestose and a nutrient component, i.e., whether or not there is a synergistic growth-promoting effect on butyric acid bacteria by the combined use of 1-kestose and a nutrient component.

[0026] Specifically, the slope of the line ab, that is, the value b / a obtained by dividing the turbidity increase at point b by the turbidity increase at point a, and the slope of the line cd, that is, the value d / c obtained by dividing the turbidity increase at point d by the turbidity increase at point c, are When (b / a) ≧ 1.0 and (d / c) / (b / a) > 1.0 is satisfied, the combination of 1-kestose and the nutritional component in question can be evaluated as having a synergistic effect on promoting the growth of butyric acid bacteria (see Figure 1B). In this case, (d / c) naturally becomes > 1.0.

[0027] The presence or absence of a countervailing effect of promoting the growth of butyric acid bacteria by using 1-kestose in combination with a nutritional component can also be evaluated in a similar manner. That is, in the example shown in FIG. 1A, point b can be replaced with an example in which the increase in turbidity is smaller than that of point a (not shown). In this case, the nutritional component has an effect of inhibiting the growth of butyric acid bacteria, and the value of b / a is smaller than 1.0, so that When (b / a) < 1.0 and (d / c) / (b / a) > 1.0 is satisfied, the combination of 1-kestose and the nutritional component can be evaluated as having a countervailing effect of promoting the growth of butyric acid bacteria (see FIG. 1C). This countervailing effect of promoting the growth of butyric acid bacteria is defined as an effect that can be exerted by combining 1-kestose with a nutritional component, as the inhibitory effect of the growth of butyric acid bacteria of the nutritional component alone is countervailed and the effect of promoting the growth of butyric acid bacteria is further countervailed, so that (d / c) > 1.0. Therefore, the countervailing effect according to the present invention can be understood as an effect that exceeds the simple countervailing effect of one inhibitory effect and the promoting effect of the other, and in a broad sense, can be included in the category of a synergistic effect.

[0028] Next, the butyric acid bacteria whose growth is promoted by the present invention are a group of bacteria capable of producing butyric acid, such as those belonging to the genus Clostridium, Faecalibacterium, Agathobacter, Roseburia, Butyrivibrio, Eubacterium, Anaerobutyricum, Anaerostipes, Coprococcus, Blautia, Ruminococcus, Butyricimonas, and Butyricicoccus.Specifically, Clostridium butyricum, Faecalibacterium duncaniae (formerly known as Faecalibacterium prausnitizii), Agathobacter rectalis (formerly known as Eubacterium rectale), Roseburia intestinalis, Roseburia faecis, Roseburia hominis, Butyrivibrio fibrisolvens, Eubacterium ramulus, Anaerobutyricum These include Anaerotipes soehngenii (formerly known as Eubacterium hallii), Anaerotipes cacae, Coprococcus catus, Coprococcus eutactus, Coprococcus comes, Blautia coccoides, Blautia hansenii, Ruminococcus gnavus, Ruminococcus faecis, Eubacterium cylindroides, etc. The butyric acid bacteria growth promoter according to the present invention promotes the growth of at least one type of bacteria, as exemplified herein. The butyric acid bacteria growth promoter according to the present embodiment exhibits a growth promoting effect on multiple types of bacteria, including at least Anaerotipes cacae, Roseburia intestinalis, and Faecalibacterium duncaniae (see Examples).

[0029] Next, 1-kestose is a fructooligosaccharide in which one molecule of fructose is bound to one molecule of sucrose, and is a trisaccharide in which fructose is bound to the fructose of sucrose via a β(2,1) glycosidic bond. It is known that 1-kestose has a growth-promoting effect on butyric acid bacteria, but the present invention represents a dramatic technological advance in that the combined use of 1-kestose and a specified nutritional component has a synergistic or countervailing growth-promoting effect on butyric acid bacteria that exceeds the effects of both on butyric acid bacteria.

[0030] 1-kestose is a known chemical substance and is available commercially, so that the commercially available products can be used as is as the formulation components according to the present embodiment. 1-kestose is also contained in vegetables, fruits, grains, etc., such as onion, garlic, banana, barley, and rye. Therefore, extracts derived from these organisms may be used as the formulation components.

[0031] As an industrial method for producing 1-kestose, a production method using a specific β-fructofuranosidase having sucrose hydrolysis activity and fructosyltransferase activity is known (JP Patent Publication 201980 / 1983, etc.). Specifically, sucrose is used as a substrate and a specific β-fructofuranosidase is reacted at 37°C to 50°C for about 20 hours to obtain an enzyme reaction solution. Since this enzyme reaction solution contains a considerable amount of 1-kestose, this sugar solution may be used as a formulation component.

[0032] Alternatively, the enzyme reaction solution may be subjected to a chromatographic separation method as disclosed in JP-A-2000-232878 to obtain a purified 1-kestose solution, which may be used as a formulation component. Alternatively, the purified solution may be concentrated and crystallized by a crystallization method as disclosed in JP-B-6-70075 to obtain 1-kestose crystals or a high-purity (e.g., purity of 98% by mass or more) 1-kestose-containing material may be used as a formulation component. Note that "purity" here means the mass % of 1-kestose in 100% by mass of the total mass of sugar. Alternatively, 1-kestose purified or further crystallized from commercially available fructooligosaccharides by the above-mentioned method may be used as a formulation component.

[0033] 1-kestose can be analyzed by a method suitable for the condition of the analytical sample among commonly known analytical methods, and a method usually selected by a person skilled in the art depending on the condition of the analytical sample can be used. In a common method, the content of 1-kestose can be analyzed by dissolving the analytical sample in water, or diluting a liquid sample with water as necessary, and subjecting it to high performance liquid chromatography (HPLC) to analyze the sugar composition. The content of 1-kestose can also be calculated by taking the ratio of the peak area in the chromatogram as the mass ratio.

[0034] Next, the nutritional components that can be combined with 1-kestose are specific nutritional components such as polyphenols, vitamins, lipids, amino acids, proteins, or minerals, and the combined use of at least one of these components with 1-kestose can provide a synergistic or countervailing effect on the growth of butyric acid bacteria. Note that the vitamins referred to here include vitamins and their derivatives as well as their salts.

[0035] Among the nutritional components, polyphenols include rutin, quercetin, catechin, hesperidin, daidzein, genistein, p-coumaric acid, caffeic acid, and vanillin. Caffeic acid may be called caffeic acid or caffeic acid, and vanillin may be called vanillin. All of these are known nutritional components and are commercially available, and products that are distributed with labels that include the names of each component, including alternative names, are at least included in the polyphenol components referred to here. In addition, each component is contained in a plant body, etc., and extracts or products thereof may be used as formulation components.

[0036] The catechins referred to here include catechin, epicatechin, gallocatechin, epigallocatechin, catechin gallate, epicatechin gallate, gallocatechin gallate, and epigallocatechin gallate, and it is sufficient to have at least one of these. When two or more types are contained, the content of catechins refers to the total amount of all the types contained.

[0037] Polyphenols may also be modified as glycosides. That is, rutin is a quercetin glycoside, and hesperidin is a flavanone glycoside. However, quercetin, daidzein, genistein, p-coumaric acid, caffeic acid, and vanillin can also exist as glycosides in plants, etc., and therefore these polyphenol components contained in extracts or products of plants, etc., may be used as formulation ingredients in the form of glycosides.

[0038] Among the nutritional components, vitamins include water-soluble vitamins such as biotin, niacin, pantothenic acid, and riboflavin, as well as derivatives of these vitamins, as well as salts of the vitamins and salts of the derivatives.

[0039] Niacin here includes nicotinic acid and nicotinamide. Also, vitamin derivatives and salts are not particularly limited as long as they are normally used in food, medicine, etc. For example, in the case of riboflavin, derivatives include riboflavin esters such as riboflavin phosphate, riboflavin acetate, riboflavin butyrate, and flavin adenine dinucleotide, and salts include riboflavin sodium phosphate, flavin sodium adenine dinucleotide, etc.

[0040] Biotin is sometimes called vitamin B7, niacin is called vitamin B3, pantothenic acid is called vitamin B5, and riboflavin is called vitamin B2 or lactoflavin. All of these are known nutrients and are commercially available, and products that are distributed with the names of each ingredient, including their alternative names, are at least included in the vitamin ingredients referred to here. There is no restriction on whether the vitamins are natural or synthetic.

[0041] Furthermore, vitamins include fat-soluble vitamins such as vitamin E and vitamin D, and include derivatives of these vitamins, as well as salts of the vitamins and salts of the derivatives.

[0042] Vitamin E includes tocopherol and tocotrienol, including α-tocopherol, β-tocopherol, γ-tocopherol, σ-tocopherol, α-tocotrienol, β-tocotrienol, γ-tocotrienol, and σ-tocotrienol. It may be either d-form or dl-form. Vitamin D includes ergocalciferol, which is vitamin D2, and cholecalciferol, which is vitamin D3. In addition, the vitamin derivatives and salts are not particularly limited as long as they are normally used in foods, medicines, etc. For example, in the case of vitamin E, derivatives such as acetate, nicotinate, and phosphate of tocopherol or tocotrienol, and in the case of vitamin D, derivatives such as sulfate of ergocalciferol or cholecalciferol, and further salts thereof, etc. are exemplified. Both vitamin E and vitamin D are known nutritional components and commercially available products are available, and products distributed with the names of each component, including their alternative names, are at least included in the vitamin components referred to here. There is no limitation as to whether the vitamins are natural or synthetic.

[0043] According to the examples described later, among the water-soluble vitamins (vitamin B group: biotin, niacin, pantothenic acid, and riboflavin), riboflavin in particular exhibited a favorable synergistic effect of promoting the growth of butyric acid bacteria. Furthermore, compared to the water-soluble vitamins, fat-soluble vitamins (α-tocopherol and cholecalciferol) exhibited a relatively more favorable countervailing effect of promoting the growth of butyric acid bacteria.

[0044] Among the nutritional components, lipids include arachidonic acid, α-linolenic acid, and docosahexaenoic acid. Arachidonic acid is a type of ω-6 fatty acid, and is a carboxylic acid consisting of a 20-carbon chain including four double bonds. On the other hand, α-linolenic acid and docosahexaenoic acid are types of ω-3 fatty acids, and α-linolenic acid is a carboxylic acid consisting of an 18-carbon chain including three cis-type double bonds. Docosahexaenoic acid is also called DNA (Docosahexaenoic acid) and is a general term for carboxylic acids having a 22-carbon chain including six double bonds. For example, the type that has cis-type double bonds at positions 4, 7, 10, 13, 16, and 19 is known as a useful nutritional component. All of these are known nutritional components and are commercially available, and products that are distributed with a labeling that includes the names of each component, including their alternative names, are at least included in the lipid components referred to here.

[0045] In addition, lipids include compounds having arachidonic acid, α-linolenic acid, and docosahexaenoic acid as constituent fatty acids. Compounds having these fatty acids as constituent fatty acids are not particularly limited as long as they are those that are normally used in foods, medicines, etc. For example, fats and oils containing arachidonic acid, α-linolenic acid, or docosahexaenoic acid as constituent fatty acids at a relatively high concentration (for example, perilla oil, linseed oil, perilla oil, etc., containing α-linolenic acid at a relatively high concentration), esters of the fatty acids, particularly triglycerides containing arachidonic acid, phospholipids, glycolipids, and other arachidonic acid salts, etc. are exemplified. Docosahexaenoic acid is contained in fish oils extracted from sardines, mackerel, horse mackerel, salmon, pacific saury, etc., fish oils derived from orbital fat of tuna, etc., and oils and oils derived from microorganisms and seaweed, and these may be used as docosahexaenoic acid in formulation components.

[0046] Among the nutritional components, the amino acid includes γ-aminobutyric acid (GABA), and the protein includes whey protein. Whey protein is a protein other than casein contained in milk-derived proteins (milk proteins). Both are known nutritional components and are commercially available, and products that are distributed with labels that include the names of each component, including their alternative names, contain at least the amino acid components or protein components referred to here.

[0047] Among the nutritional components, minerals include potassium, magnesium, and iron. These minerals may be contained in the liquid preparation as ions. The mineral source used as the preparation component is not particularly limited as long as it is acceptable for incorporation into food or medicine. Examples of the potassium source include potassium lactate, potassium carbonate, potassium chloride, potassium phosphate (potassium dihydrogen phosphate, dipotassium hydrogen phosphate, tripotassium phosphate), potassium gluconate, potassium ascorbate, potassium L-glutamate, etc. Examples of the magnesium source include magnesium chloride, magnesium silicate, magnesium oxide, magnesium hydroxide, magnesium carbonate, magnesium sulfate, trimagnesium phosphate, magnesium stearate, magnesium L-glutamate, etc. Examples of the iron source include ferrous or ferric iron such as iron citrate, iron sulfide, iron sulfate, iron chloride, iron hydrochloride, iron gluconate, and iron lactate, and sodium iron chlorophyllin. All of these minerals (mineral sources) are known nutritional components and are commercially available, and products distributed under the labeling including the names of each component including their alternative names are at least included in the mineral components referred to here.

[0048] The nutritional components according to the present embodiment include, in addition to those specifically mentioned, derivatives of each component, salts of each component and salts of derivatives, intermediates, precursors, and other compounds containing each component, within the scope that can be understood by a person skilled in the art to be formulated for the purpose of intake (administration) of the nutritional components. In addition, each nutritional component can be analyzed by a method suitable for the condition of the analysis sample among commonly known analytical methods, and a method usually selected by a person skilled in the art according to the condition of the analysis sample, such as HPLC, GC-MS, FT-IR, etc., can be appropriately used.

[0049] The butyric acid bacteria growth promoter according to the present embodiment (hereinafter, the present agent) is not limited to ingredients other than 1-kestose and the specified nutritional components specified as contained ingredients, and may contain other nutritional components (sugars, dietary fiber, polyphenols, vitamins, vitamin-like substances, lipids, amino acids, proteins, minerals, etc.), other food ingredients, medicinal ingredients, herbal medicines, etc. The present agent can be provided as a prebiotic containing 1-kestose and the specified nutritional components. By ingesting (administering) this, the growth of butyric acid bacteria normally present in the intestine can be promoted. Alternatively, by adding butyric acid bacteria as probiotics to 1-kestose and the specified nutritional components as prebiotics, the present agent can be provided as a synbiotic containing 1-kestose, the specified nutritional components, and butyric acid bacteria. By ingesting (administering) this, the growth of the ingested (administered) butyric acid bacteria and the butyric acid bacteria normally present in the intestine can be promoted.

[0050] In addition, the dosage form of the present agent is not limited, and for example, it can be appropriately formed into oral solid preparations (including supplements) such as tablets, granules, powders, and capsules, oral liquid preparations such as oral liquids and syrups, and parenteral liquid preparations such as injections. In addition, an appropriate drug delivery system (DDS) may be applied. It can be appropriately formed and formulated into each dosage form by known methods, and additives such as excipients, binders, lubricants, colorants, disintegrants, thickeners, preservatives, stabilizers, and pH adjusters that are usually added during forming and formulation may be included in the present agent. In addition, the administration method of the present agent is not limited, and oral administration, sublingual administration, administration from a pharyngeal fistula, a gastrostomy, or an intestinal fistula, etc. can be appropriately selected. In addition, it may be administered once or multiple times per day.

[0051] In addition, the agent itself can be applied as a food, a medicine, etc. Alternatively, the agent can be applied as a food composition, a medicine composition, etc. That is, when the agent is applied as a composition, it can take the form of a food, a medicine, etc. containing the agent as a composition. In other words, it can take the form of a food, a medicine, etc. containing the agent as a composition. The term "medicine" as used herein means the medicines specified in Article 2, paragraph 1 of the Law on Ensuring Quality, Efficacy and Safety of Pharmaceuticals, Medical Devices, etc., excluding the parentheses at the end of paragraphs 2 and 3 of the same paragraph, i.e., including quasi-drugs. The term "food" as used herein means anything that is eaten or drunk, including beverages.

[0052] When this agent is applied as a food or food composition, the food which is this agent or the food which contains this agent can be applied as a food to be taken for the purpose of promoting the growth of butyric acid bacteria and the health function equivalent thereto. In other words, it may be applicable as a product which displays or claims a specific health function as a food for specified health use under the Health Promotion Act or a food with functional claims under the Food Labeling Act.

[0053] In addition, the form of the product containing the agent as a composition is not limited. That is, the food, medicine, etc. containing the agent as a composition may also be in the form of oral solid preparations (including supplements) such as tablets, granules, powders, capsules, etc., oral liquid preparations such as oral liquids and syrups, and parenteral liquid preparations such as injections, like the agent. Therefore, a preparation containing 1-kestose and a nutritional component may be a food, medicine, etc. containing the agent as a composition, as well as a food, medicine, etc. containing the agent as a composition.

[0054] Furthermore, 1-kestose is also contained in foods that humans ingest on a daily basis, is a substance that has been consumed since ancient times, and has not been found to be toxic or mutagenic in toxicity tests and genotoxicity tests, so it can be used as a food ingredient with high safety (Food and Development, December 2014, Vol. 49, No. 12, p. 8-10). Therefore, the present agent may contain 1-kestose and may be ingested (administered) in an amount exceeding the amount at which the growth-promoting effect of butyric acid bacteria was confirmed in the examples described below. On the other hand, nutritional components with a tolerable upper limit must be ingested (administered) within a range not exceeding the tolerable upper limit, but in the examples described below, any of the nutritional components according to this embodiment exhibited a growth-promoting effect on butyric acid bacteria when combined with 1-kestose within a range not exceeding the tolerable upper limit for adults.

[0055] In addition, 1-kestose has extremely low hygroscopicity and excellent solubility and heat resistance, making it highly convenient for food processing, and has a sweetness similar to that of sugar, so it can be used as a sugar substitute. Taking advantage of these properties of 1-kestose, foods containing the agent as a composition can be suitably applied as various foods, such as gums, gummies, candies and other confectioneries, flake- or puff-shaped cereals, bars, jellies, drink jellies, swallowable foods, beverages, etc., containing 1-kestose and a specified nutritional component. EXAMPLES

[0056] A culture test of butyric acid bacteria was conducted in a system containing 1-kestose and specified nutritional components. The following 1-kestose and nutritional components were used in the test (note that product names will not be listed for products that are sold under the names of the components or reagents themselves). 1-Kestose (iKes Crystal (registered trademark) (purity 95% by mass), Itochu Sugar Co., Ltd.) Polyphenols: Rutin (Tokyo Chemical Industry Co., Ltd.), Quercetin (Tokyo Chemical Industry Co., Ltd.), Catechin (Tokyo Chemical Industry Co., Ltd.), Hesperidin (Fujifilm Wako Pure Chemical Industries Co., Ltd.), Daidzein (Fujifilm Wako Pure Chemical Industries Co., Ltd.), Genistein (Fujifilm Wako Pure Chemical Industries Co., Ltd.), p-Coumaric acid (Fujifilm Wako Pure Chemical Industries Co., Ltd.), Caffeic acid (Tokyo Chemical Industry Co., Ltd.), Vanillin (Tokyo Chemical Industry Co., Ltd.) Vitamins: Biotin (FUJIFILM Wako Pure Chemical Industries, Ltd.), Niacin (FUJIFILM Wako Pure Chemical Industries, Ltd.), Pantothenic Acid (FUJIFILM Wako Pure Chemical Industries, Ltd.), Riboflavin (FUJIFILM Wako Pure Chemical Industries, Ltd.), α-Tocopherol (FUJIFILM Wako Pure Chemical Industries, Ltd.), Cholecalciferol (FUJIFILM Wako Pure Chemical Industries, Ltd.) Lipids: Arachidonic acid (Fujifilm Wako Pure Chemical Industries, Ltd.), α-linolenic acid (Fujifilm Wako Pure Chemical Industries, Ltd.), docosahexaenoic acid (Fujifilm Wako Pure Chemical Industries, Ltd.) Amine acids, proteins: γ-aminobutyric acid (FUJIFILM Wako Pure Chemical Industries, Ltd.), whey protein (THERMAX 690, Nissei Kyoeki Co., Ltd.) Minerals: Dipotassium hydrogen phosphate K2HPO4 (FUJIFILM Wako Pure Chemical Industries, Ltd.), Magnesium sulfate MgSO4·7H2O (FUJIFILM Wako Pure Chemical Industries, Ltd.), Iron(III) chloride FeCl3·6H2O (FUJIFILM Wako Pure Chemical Industries, Ltd.)

[0057] The basal medium, YCFA medium, was prepared as follows: 10 g of tryptone (Nihon Seiyaku Co., Ltd.), 2.5 g of yeast extract (Nippon Becton Dickinson Co., Ltd.), 4.0 g of NaHCO3 (Fujifilm Wako Pure Chemical Industries, Ltd.), 1.0 g of L-cysteine ​​hydrochloride hydrate (Fujifilm Wako Pure Chemical Industries, Ltd.), 1 mL of 1 mg / L resazurin sodium salt (Tokyo Chemical Industry Co., Ltd.) prepared with distilled water, 150 mL each of mineral solutions I and II, 6.2 mL of VFA mix, and 10.0 mL of hemin solution were dissolved in 660 mL of distilled water, the pH was adjusted to 7.45, residual oxygen was replaced with nitrogen gas, and the mixture was sterilized in an autoclave at 121°C for 15 minutes. 1.0 mL each of filter-sterilized vitamin solutions I and II were added to make a total of 1 L to prepare YCFA medium.

[0058] Among the above YCFA medium compositions, mineral solution I was prepared by dissolving 3.0 g of K2HPO4 (FUJIFILM Wako Pure Chemical Industries, Ltd.) in 1.0 L of distilled water. Mineral solution II was prepared by dissolving 3.0 g of K2HPO4 (FUJIFILM Wako Pure Chemical Industries, Ltd.), 6.0 g of (NH4)2SO4 (FUJIFILM Wako Pure Chemical Industries, Ltd.), 6.0 g of NaCl (FUJIFILM Wako Pure Chemical Industries, Ltd.), 0.6 g of MgSO4·7H2O (FUJIFILM Wako Pure Chemical Industries, Ltd.), and 0.6 g of CaCl2·2H2O (FUJIFILM Wako Pure Chemical Industries, Ltd.) in 1.0 L of distilled water. The VFA mix was prepared by mixing 17.0 mL of acetic acid (Tokyo Chemical Industry Co., Ltd.), 6.0 mL of propionic acid (Tokyo Chemical Industry Co., Ltd.), 1.0 mL of n-valeric acid (Fujifilm Wako Pure Chemical Industries Co., Ltd.), 1.0 mL of isovaleric acid (Fujifilm Wako Pure Chemical Industries Co., Ltd.), 1.0 mL of isobutyric acid (Fujifilm Wako Pure Chemical Industries Co., Ltd.), and 26 mL of 10N NaOH (Fujifilm Wako Pure Chemical Industries Co., Ltd.). The hemin solution was prepared by dissolving 0.28 g of KOH (Fujifilm Wako Pure Chemical Industries Co., Ltd.), 25.0 mL of ethanol (Nacalai Tesque, Inc.), and 0.1 g of hemin (Fujifilm Wako Pure Chemical Industries Co., Ltd.) in 100 mL of distilled water. Vitamin solution I was prepared by dissolving 5.0 mg of biotin (FUJIFILM Wako Pure Chemical Industries, Ltd.), 5.0 mg of vitamin B12 (FUJIFILM Wako Pure Chemical Industries, Ltd.), 15.0 mg of p-aminobenzoic acid (FUJIFILM Wako Pure Chemical Industries, Ltd.), 25.0 mg of folic acid (FUJIFILM Wako Pure Chemical Industries, Ltd.), and 75.0 mg of pyridoxine hydrochloride (FUJIFILM Wako Pure Chemical Industries, Ltd.) in 500 mL of distilled water. Vitamin solution II was prepared by dissolving 25.0 mg of thiamine hydrochloride (FUJIFILM Wako Pure Chemical Industries, Ltd.) and 25.0 mg of riboflavin (FUJIFILM Wako Pure Chemical Industries, Ltd.) in 500 mL of distilled water.

[0059] A 96-deep well plate was filled with 500 μL of YCFA medium, and each of the main agent 1-kestose and each nutrient component as a co-agent, which were serially diluted, and an appropriate amount of water was added to a total of 1 mL to obtain the concentrations shown in Table 1 to prepare [Kes + nutrient component-added medium] (verification medium d). In addition, the nutrient component (solution) portion of verification medium d was replaced with water [Kes-added medium] (verification medium c), the 1-kestose (solution) portion of verification medium d was replaced with water [nutrient component-added medium] (verification medium b), and an appropriate amount of water was added to 500 μL of YCFA medium to make a total of 1 mL [supplement-free medium] (verification medium a). As a result, the following four test sections were set up. Test group a (verification medium a): YCFA medium + water (supplement-free medium) Test group b (verification medium b): YCFA medium + nutrients + water (nutrient-supplemented medium) Test group c (verification medium c): YCFA medium + 1-kestose + water (Kes-supplemented medium) Test group d (verification medium d): YCFA medium + 1-kestose + nutrients + water (Kes + nutrients added medium)

[0060] [Table 1]

[0061] Anaerotipes cacae (JCM 13470), Roseburia intestinalis (JCM 17583), and Faecalibacterium duncaniae (JCM 31915) provided by RIKEN BRC-JCM were used as the culture subjects for verifying the growth promotion of butyric acid bacteria. YCFA medium containing 0.5 mass% glucose as a carbon source was used as the pre-culture medium, and after culturing the three types of bacteria for 16 to 24 hours, 20 μL of each of the verification media (2 mass% per mL of medium) was inoculated and cultured.

[0062] The amount of butyric acid bacteria proliferation was measured and calculated 24 hours and 48 hours after the start of the culture. Specifically, the amount of turbidity increase, which represents the net turbidity calculated by subtracting the turbidity measurement value at the start of the culture as a blank value from the turbidity measurement value of each test medium at 24 hours and 48 hours after the start of the culture, was used as the amount of butyric acid bacteria proliferation in each test medium at that time. The turbidity was measured using a plate reader (Sunrise Rainbow, Tecan) at an absorbance wavelength of 660 nm. The turbidity at the start of the culture was measured for each test medium before inoculation.

[0063] Based on the calculated amount of growth of butyric acid bacteria, the growth promotion effect of butyric acid bacteria was evaluated. The growth promotion effect was evaluated according to the above-mentioned provisions of the main text. [(growth-promoting or growth-inhibiting effect when Kes is used in combination with a nutritional component) / (growth-promoting or growth-inhibiting effect when only a nutritional component is added)] The formula (d / c) / (b / a) was calculated (here, d represents the amount of growth in test plot d (verification medium d), c represents the amount of growth in test plot c (verification medium c), b represents the amount of growth in test plot b (verification medium b), and a represents the amount of growth in test plot a (verification medium a)). When (b / a) ≧ 1.00, it was evaluated that (d / c) / (b / a) > 1.00 was satisfied, and when (d / c) > 1.00, it was evaluated that there was a synergistic effect of promoting the growth of butyric acid bacteria. In addition, when (b / a) < 1.00, (d / c) / (b / a) > 1.00 is satisfied, and when (d / c) > 1.00, it is evaluated as a countervailing effect on the growth of butyric acid bacteria. Furthermore, the degree of synergistic or countervailing effect of promoting the growth of butyric acid bacteria was classified according to the following criteria. If (d / c) / (b / a) = 1.00 or less: no effect (blank) = Above 1.00 and below 1.05: Effective (+) = 1.05 or more but less than 1.10: Effective (++) = 1.10 or more and less than 1.20: Effective (+++) = 1.20 or more and less than 2.00: Effective (*) = 2.00 or more: Effective (**) The results are shown in Tables 2 to 6.

[0064] [Table 2]

[0065] As shown in Table 2, the combined use of rutin at a ratio of 100 μg / mL, quercetin at a ratio of 2 μg / mL, catechin at a ratio of 2 μg / mL or more, hesperidin at a ratio of 2 μg / mL or 100 μg / mL, daidzein at a ratio of 2 μg / mL or more, genistein at a ratio of 2 μg / mL or more, p-coumaric acid at a ratio of 100 μg / mL, caffeic acid at a ratio of 20 μg / mL, and vanillin at a ratio of 2 μg / mL to 20 μg / mL with respect to 1-kestose 5 mg / mL exhibited a synergistic or countervailing effect on the growth of butyric acid bacteria. In particular, isoflavones (daidzein, genistein) exhibited a favorable countervailing effect on the growth of butyric acid bacteria.

[0066] [Table 3]

[0067] As shown in Table 3, the combination of biotin at a ratio of 1 μg / mL, niacin at a ratio of 1.7 μg / mL, pantothenic acid at a ratio of 20 μg / mL or more, riboflavin at a ratio of 2 μg / mL or more, α-tocopherol at a ratio of 2 μg / mL or more, and cholecalciferol at a ratio of 2 μg / mL to 20 μg / mL with 5 mg / mL of 1-kestose each exhibited synergistic or countervailing effects on the growth of butyric acid bacteria.

[0068] Among the water-soluble vitamins (vitamin B group: biotin, niacin, pantothenic acid, and riboflavin), riboflavin in particular exerted a favorable synergistic effect on the growth of butyric acid bacteria. In addition, compared to the water-soluble vitamins, the fat-soluble vitamins (α-tocopherol and cholecalciferol) exerted a relatively more favorable countervailing effect on the growth of butyric acid bacteria.

[0069] [Table 4]

[0070] As shown in Table 4, the combined use of arachidonic acid at a ratio of 20 μg / mL or more, α-linolenic acid at a ratio of 2 μg / mL or more, and docosahexaenoic acid at a ratio of 2 μg / mL or more with 1-kestose 5 mg / mL each exhibited a countervailing effect on the growth of butyric acid bacteria. In particular, docosahexaenoic acid exhibited a favorable countervailing effect on the growth of butyric acid bacteria.

[0071] [Table 5]

[0072] As shown in Table 5, the combined use of 5 mg / mL 1-kestose with γ-aminobutyric acid at a ratio of 1000 μg / mL or more and with whey protein at a ratio of 500 μg / mL or more exhibited synergistic or countervailing growth-promoting effects on butyric acid bacteria. In particular, whey protein exhibited a favorable countervailing growth-promoting effect on butyric acid bacteria.

[0073] [Table 6]

[0074] As shown in Table 6, the combination of dipotassium hydrogen phosphate at a ratio of 6 mg / mL, magnesium sulfate at a ratio of 2.3 mg / mL or more, and iron(III) chloride at a ratio of 0.006 mg / mL or more with 1-kestose at 5 mg / mL each exerted synergistic or countervailing effects on the growth of butyric acid bacteria.

[0075] Although the data supporting the present invention is shown here, nutritional components other than those shown here were also tested. These did not necessarily exhibit a synergistic or countervailing effect on promoting the growth of butyric acid bacteria, which shows that the present invention (the invention related to the claims) is a combination that is the result of extensive research based on the verification of various nutritional components.

Claims

1. A synergistic or counteractive growth promoter for butyric acid bacteria, comprising 1-kestose and a nutrient component, The butyric acid bacteria growth promoter, wherein the nutritional component is one or more polyphenols selected from rutin, quercetin, catechin, hesperidin, daidzein, genistein, p-coumaric acid, caffeic acid, and vanillin.

2. A synergistic or counteractive growth promoter for butyric acid bacteria, comprising 1-kestose and a nutrient component, The butyric acid bacteria growth promoter, wherein the nutritional component is one or more vitamins selected from biotin, niacin, pantothenic acid, riboflavin, vitamin E, and vitamin D, derivatives of these vitamins, salts of the vitamins, and salts of the derivatives.

3. A synergistic or counteractive growth promoter for butyric acid bacteria, comprising 1-kestose and a nutrient component, The growth promoter for butyric acid bacteria, wherein the nutritional component is one or more lipids selected from arachidonic acid, α-linolenic acid, docosahexaenoic acid, and compounds having these fatty acids as constituent fatty acids.

4. A synergistic or counteractive growth promoter for butyric acid bacteria, comprising 1-kestose and a nutrient component, The butyric acid bacteria growth promoter, wherein the nutritional component is one or more amino acids or proteins selected from γ-aminobutyric acid and whey protein.

5. A synergistic or counteractive growth promoter for butyric acid bacteria, comprising 1-kestose and a nutrient component, A butyric acid bacteria growth promoter, wherein the nutritional component is one or more minerals selected from potassium, magnesium, and iron.

6. The butyric acid bacteria growth promoter according to claim 2, wherein said vitamin E is α-tocopherol and said vitamin D is cholecalciferol.

7. The butyric acid bacteria growth promoter according to any one of claims 1 to 6, which is a synbiotic containing butyric acid bacteria in addition to the 1-kestose and the nutritional components.

8. A food or medicine comprising the butyric acid bacteria growth promoter according to any one of claims 1 to 6.

Images