Composition for promoting growth of agathobacter bacteria

Abstract

To provide a composition for promoting growth of Agathobacter bacteria in the intestine, a food product, medicine, and feed each including the composition.SOLUTION: A composition for promoting growth of Agathobacter bacteria comprises, as an active ingredient, a bacterial cell or culture belonging to Lactobacillus paragasseri. In particular, in the composition for promoting growth of Agathobacter bacteria, Lactobacillus paragasseri is Lactobacillus paragasseri SBT2055 (FERM BP-10953).SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to a composition for promoting the growth of Agasobacter bacteria, which contains Lactobacillus paragaceri cells and / or a culture as an active ingredient. [Background technology]

[0002] The human intestine is home to 100 trillion bacteria of 1,000 different species, forming a complex symbiotic relationship known as the intestinal microbiota. In addition to forming a symbiotic relationship with other bacteria, intestinal bacteria also exert various physiological effects on the host via the intestinal wall, and it has become clear in recent years that they play an important role in maintaining health and preventing disease, and their association with gastrointestinal diseases such as ulcerative colitis and chronic constipation, cardiovascular disease, and cancer has been pointed out.

[0003] Intestinal bacteria metabolize indigestible dietary fiber and oligosaccharides to produce short-chain fatty acids such as acetic acid, propionic acid, and butyric acid. One of these, butyric acid, not only inhibits the growth of harmful bacteria by maintaining an acidic intestine, but is also used as an energy source for intestinal epithelial cells and is associated with maintaining homeostasis of the intestinal immune system, including biological defense and anti-inflammatory effects. Therefore, intestinal bacteria that produce butyric acid have attracted particular attention.

[0004] In particular, the occupancy rate of Agasobacter bacteria, a type of enterobacteria that produces butyric acid, in the intestines of patients with ulcerative colitis, anorexia nervosa, depression, etc. is significantly decreased, suggesting a relationship with these diseases (Non-Patent Documents 1 to 3). In addition, it has been reported that, in a test using a model mouse for ulcerative colitis, an increase in Agasobacter bacteria in the intestines was observed as symptoms were alleviated (Non-Patent Document 4). In light of this background, Agasobacter bacteria may be expected to be useful probiotics for the treatment of certain diseases.

[0005] Since Agasobacter bacteria are obligate anaerobes, there are many challenges in terms of production and distribution for practical use as food, medicine, or feed for oral ingestion. On the other hand, the composition of the intestinal flora can change due to the influence of various factors such as diet and living environment. There have been many reports that the ingestion of compositions containing probiotics and prebiotics promotes the proliferation of specific intestinal bacteria and improves the intestinal environment.

[0006] Previous studies have reported that oral ingestion of the lactic acid bacterium Lactobacillus plantarum in colitis-induced mice increases the occupancy rate of Agasobacter bacteria in the fecal flora (Non-Patent Document 4). It has also been reported that the abundance of desirable bacterial species was significantly increased in COVID-19 patients who orally ingested a synbiotic containing a prebiotic composition containing xylooligosaccharides, galactooligosaccharides, and corn dietary fiber, and a probiotic composition containing Bifidobacterium adolescentis, Bifidobacterium bifidum, and Bifidobacterium longum, compared with the control group (Patent Document 1).

[0007] In these examples, the oral ingestion of some species of lactic acid bacteria as food has been found to promote the growth of Agasobacter bacteria in the intestines, but the effects of other species have not been clarified. In addition, there is no mention of their use as food for the purpose of disease prevention in healthy humans. Therefore, the objective of the present invention is to find a composition such as a probiotic or prebiotic that promotes the growth of Agasobacter bacteria in the intestines when orally ingested, and to provide a food or drink, a pharmaceutical product, and a feed for promoting the growth of Agasobacter bacteria that contain the composition. [Prior art documents] [Patent documents]

[0008] [Patent Document 1] Special Publication No. 2023-523470 [Non-patent literature]

[0009] [Non-Patent Document 1] Inflamm Bowel Dis. 2013:19:481-8 [Non-Patent Document 2] Gut Microbes. 2021:13:e1902771 [Non-Patent Document 3] Nutrients. 2023:15:1931 [Non-Patent Document 4] Food Funct. 2023:14:3982-93 Summary of the Invention [Problem to be solved by the invention]

[0010] An objective of the present invention is to provide a composition for promoting the growth of Agathobacter bacteria, and a food or beverage product for promoting the growth of Agathobacter bacteria, a pharmaceutical product for promoting the growth of Agathobacter bacteria, and a feed product for promoting the growth of Agathobacter bacteria, which contain the composition. [Means for solving the problem]

[0011] The present inventors have found that oral ingestion of a composition containing a bacterial cell and / or a culture of Lactobacillus paragaceri increases the number of Agasobacter bacteria in the intestine, and have completed the present invention. <1> A composition for promoting the growth of bacteria of the genus Agassobacter, comprising as an active ingredient a bacterial cell and / or a culture product of the species Lactobacillus paragasseri. <2> The Lactobacillus paragaceri is Lactobacillus paragaceri SBT2055 (FERM BP-10953); <1> A composition for promoting the growth of Agasobacter bacteria as described above. <3> The promotion of Agasobacter proliferation is due to an increase in the occupancy rate of Agasobacter in the human intestinal flora. <1> or <2> A composition for promoting the growth of Agasobacter bacteria as described above. <4> <1> or <2> 2. A food or drink for promoting the growth of Agasobacter bacteria, comprising the composition for promoting the growth of Agasobacter bacteria described in claim 1. <5> <1> or <2> 2. A pharmaceutical composition for promoting the growth of Agasobacter bacteria, comprising the composition for promoting the growth of Agasobacter bacteria described in claim 1. <6> <1> or <2> 2. A feed for promoting the growth of bacteria of the genus Agasobacter, comprising the composition for promoting the growth of bacteria of the genus Agasobacter described above. Effect of the Invention

[0012] According to the present invention, a composition for promoting the growth of bacteria of the genus Agasobacter containing a bacterium and / or a culture of Lactobacillus paragaceri as an active ingredient can be provided.Furthermore, a food for promoting the growth of bacteria of the genus Agasobacter, a pharmaceutical product for promoting the growth of bacteria of the genus Agasobacter, and a feed for promoting the growth of bacteria of the genus Agasobacter, each containing the composition, can be provided. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0013] DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS A specific embodiment of the composition for promoting the growth of Agasobacter bacteria according to the present invention will be described in detail below.

[0014] In the present invention, the composition for promoting the growth of Agathobacter bacteria means a composition capable of increasing Agathobacter bacteria. In addition, examples of the increase in Agathobacter bacteria include an increase in the occupancy rate of Agathobacter bacteria in a specific bacterial flora such as in the intestine, and an increase in the absolute amount of Agathobacter bacteria. Examples of species of bacteria belonging to the Agathobacter genus include Agathobacter rectalis, Agathobacter ruminis, and Agathobacter faecis.

[0015] (Lactobacillus paragaceri) The present invention uses Lactobacillus paragasseri as an active ingredient, which is a bacterium that exhibits an effect of promoting the growth of Agasobacter bacteria. Preferred examples thereof include Lactobacillus paragaceri SBT2055 (FERM BP-10953) and strains substantially equivalent to Lactobacillus paragaceri SBT2055 (FERM BP-10953). The substantially equivalent strain is one that exhibits an effect of promoting the proliferation of bacteria of the genus Agasobacter, has a base sequence of its 16S rRNA gene that is 98% or more, preferably 99% or more, more preferably 100% homologous to the base sequence of the 16S rRNA gene of Lactobacillus paragaceri SBT2055, and preferably has the same bacteriological properties as Lactobacillus paragaceri SBT2055. Furthermore, as long as the effects of the present invention are not impaired, the strain may be one that has been bred from Lactobacillus paragaceri SBT2055 or a strain substantially equivalent thereto by mutation treatment, genetic recombination, selection of natural mutants, or the like. Lactobacillus paragaceri SBT2055 is a strain whose species was reclassified as a result of a revision of taxonomy, and is the same as the strain described as Lactobacillus gasseri SBT2055 in previous literature. In addition, the Lactobacillus paragaceri strains exhibiting the effect of promoting the proliferation of Agasobacter bacteria used in the present invention can also be obtained by screening for strains exhibiting the effect of promoting the proliferation of Agasobacter bacteria under desired conditions, such as the intestinal flora, using the evaluation method described below.

[0016] (Preparation of active ingredient of composition for promoting growth of Agasobacter bacteria) The active ingredient of the present invention is the above-mentioned Lactobacillus paragaceri bacteria and / or bacteria culture. The Lactobacillus paragaceri bacteria can be obtained by culturing Lactobacillus paragaceri according to a conventional method for culturing lactic acid bacteria and isolating the bacteria from the resulting culture by a collection means such as centrifugation. The composition containing Lactobacillus paragaceri can be prepared using not only purely isolated bacteria, but also bacteria-containing substances such as cultures and suspensions, as well as dried products, concentrates, pastes, etc. of these. The bacteria may be live or dead bacteria, and are preferably live bacteria. In addition, examples of the culture of Lactobacillus paragaceri include a culture obtained by culturing Lactobacillus paragaceri in a general lactic acid bacteria medium and a culture supernatant containing metabolic products produced by the bacteria during the culture. In addition, dairy products such as cheese, fermented milk, and lactic acid bacteria drinks prepared with Lactobacillus paragaceri are also included in the bacterial cells or cultures of the present invention. Furthermore, Lactobacillus paragaceri may be used alone or in combination of two or more types. Furthermore, the bacterial cells and bacterial cell cultures may be used in combination.

[0017] (Composition for promoting the growth of Agasobacter bacteria) The composition for promoting the growth of Agasobacter bacteria of the present invention may contain only the active ingredient Lactobacillus paragaceri cells and / or cultures, and may contain only the active ingredient or may contain other ingredients within a range that does not interfere with the growth promotion effect of Agasobacter bacteria. The composition for promoting the growth of Agasobacter bacteria of the present invention is preferably for oral use, and the other ingredients include food and drink compositions, oral pharmaceutical compositions, feed compositions, etc. that are suitable for oral use. Furthermore, it may contain probiotics such as other lactic acid bacteria within a range that does not interfere with the growth promotion of Agasobacter bacteria.

[0018] (Food and drink for promoting the growth of Agasobacter bacteria) The form of a food or drink which is one embodiment of the composition for promoting the growth of Agasobacter bacteria of the present invention will be described below. The food or drink for promoting the growth of bacteria of the genus Agathobacter of the present invention may be in any form as long as it does not impair the effect of promoting the growth of bacteria of the genus Agathobacter. Examples of the form of food and beverage products for promoting the growth of Agasobacter bacteria of the present invention include Lactobacillus paragaceri bacteria, cultures containing the bacteria, and fermented foods such as fermented milk and cheese prepared using lactic acid bacteria containing Lactobacillus paragaceri, as well as processed foods such as bread, snacks, cakes, pudding drinks, noodles, and sausages containing the above-mentioned bacteria, cultures containing the bacteria, fermented foods such as fermented milk and cheese, and various types of powdered milk, infant foods, supplements, etc.

[0019] The classification of the above-mentioned food and drink is not particularly limited, and the food and drink can be general food, health functional food, special purpose food, etc. In addition, the health functional food can be a specific health food, a nutrient functional food, a functional food, etc.

[0020] When indicating the use of the composition for promoting the growth of Agasobacter bacteria, indications may be given as stipulated by various laws, enforcement regulations, guidelines, etc. Indications of use may include indications on packages such as wrapping and containers, as well as indications on advertising media such as pamphlets.

[0021] (Drugs for promoting the growth of Agasobacter bacteria, animal feed for promoting the growth of Agasobacter bacteria) The form of a pharmaceutical or animal feed, which is one embodiment of the composition for promoting the growth of Agasobacter bacteria of the present invention, will be described below. When preparing pharmaceuticals, excipients, stabilizers, flavoring agents, etc. permitted for pharmaceutical use can be appropriately mixed and used during formulation, and excipients, binders, disintegrants, lubricants, flavoring agents, suspending agents, coating agents, and other optional drugs can also be mixed and formulated within a range that does not impair the effects of the present invention. Dosage forms that can be used include tablets, capsules, granules, powders, powders, syrups, etc., and these are administered orally. When included in animal feed, the same effects can be expected from animals that ingest it.

[0022] (Intake amount) In order to exert the effect of promoting the proliferation of bacteria of the genus Agasobacter according to the present invention, it is desirable for adults to ingest 1 to 1,000 mg / day in terms of lactic acid bacteria body weight and 100 to 100,000 mg / day in terms of culture weight. When preparing foods and beverages for promoting the proliferation of Agasobacter bacteria containing Lactobacillus paragaceri of the present invention, the content of Lactobacillus paragaceri is not particularly limited, and may be appropriately adjusted according to ease of production, a preferred daily dose, etc. For example, when the form is liquid, it is 1×10 5 cells / ml ~ 1 × 10 10 The preferred concentration is 1×10 cells / ml. 5 cells / g ~ 1 × 10 10 It is preferable to use cells / g.

[0023] (Evaluation method) The effect of the present invention in promoting the growth of bacteria of the genus Agathobacter can be evaluated by, for example, an increase in the occupancy rate of bacteria of the genus Agathobacter or an increase in the bacterial cell mass of bacteria of the genus Agathobacter under specific conditions. The former can be evaluated, for example, by the increase in the occupancy rate of Agasobacter bacteria in the intestines of a group of humans or animals that have orally ingested a target composition for a certain period of time. As for the increase in the occupancy rate of Agasobacter bacteria in the intestines, a stool sample is collected and a comprehensive analysis of the intestinal flora is performed. If the occupancy rate of Agasobacter bacteria in the feces is higher than that of a group that has not ingested the composition or at the start of ingestion, it can be evaluated that the composition has an effect of promoting the increase of Agasobacter bacteria in the intestines. In the latter case, for example, when a target composition is added to a culture medium for culturing Agasobacter bacteria and the increase in the bacterial cell mass of Agasobacter bacteria is promoted compared to a medium without the addition of the composition, it can be evaluated that the target active ingredient has an effect of promoting the increase of Agasobacter bacteria.

[0024] (Administration, Ingestion Target) The subject of administration or ingestion of the composition for promoting the increase of Agasobacter bacteria of the present invention is a human who needs an increase in Agasobacter bacteria in the intestine. Since it has been reported that Agasobacter bacteria are reduced in patients with ulcerative colitis, anorexia nervosa, depression, etc., administration of the composition of the present invention to these patients is expected to increase Agasobacter bacteria in the intestine and improve the symptoms of the disease. It has also been confirmed that the composition of the present invention promotes the increase of Agasobacter bacteria even when administered to healthy humans. Therefore, it is expected to prevent the above-mentioned disease. Furthermore, since the increase of Agasobacter bacteria is promoted even when administered to humans who tend to be constipated, it is expected to improve or prevent the above-mentioned symptoms. In addition, animals other than humans in a similar condition can also be administered or ingested. EXAMPLES

[0025] In the following, examples will be given in order to more specifically explain the configuration and effects of the present invention, but the present invention is not limited to these examples.

[0026] [Example 1] Study on the effect of Lactobacillus paragaceri intake on Agasobacter bacteria in humans (1) 1. Test meal The test food used was a capsule food containing Lactobacillus paragaceri SBT2055 (product name: Lactobacillus gasseri SP strain capsule, manufactured by Megmilk Snow Brand Co., Ltd.). The capsule contains Lactobacillus paragaceri SBT2055 bacteria and starch, and by taking three capsules, more than one billion Lactobacillus paragaceri SBT2055 bacteria can be ingested. In addition, a capsule in which the Lactobacillus paragaceri SBT2055 bacteria in the capsule was replaced with starch was manufactured and used as a placebo.

[0027] 2. Test Method 72 healthy adult men and women (36 in the test diet group and 36 in the placebo group) with a weekly bowel movement frequency of 3-5 times were selected and orally administered 3 capsules per day for 4 weeks. Stool samples were collected before and 4 weeks after the start of the study, and the stool samples were collected in dedicated containers and promptly collected under refrigerated and anaerobic conditions. A comprehensive analysis of the intestinal flora was performed using the collected stool samples, and the bacterial occupancy rate of Agasobacter bacteria in the total number of eubacteria in the feces was measured. The comprehensive analysis of the intestinal flora was performed by 16S rRNA amplicon sequencing analysis using Quantitative Insights Into Microbial Ecology 2 (Qiime2), and the database used was SILVA 138.

[0028] 3. Test Results The percentage of Agasobacter bacteria in the total number of eubacteria in the feces of the subjects before and after the start of the study is shown in Table 1. The unit is %.

[0029] [Table 1]

[0030] As shown in Table 1, the intestinal occupancy rate of Agasobacter bacteria in the test food group was significantly higher than that of the placebo group four weeks after the start of the study. The intestinal occupancy rate was also confirmed to have increased compared to before the start of the study. Therefore, it was found that oral ingestion of Lactobacillus paragaceri bacteria promotes the increase of Agasobacter bacteria in healthy humans who tend to suffer from constipation.

[0031] [Example 2] Study on the effect of Lactobacillus paragaceri intake on Agasobacter bacteria in humans (2) 1. Test meal As in Example 1, a capsule food containing Lactobacillus paragaceri SBT2055 (product name: Lactobacillus gasseri SP strain capsule, manufactured by Megmilk Snow Brand Co., Ltd.) was used as the test food.

[0032] 2. Test Method 95 healthy adult men and women were selected and orally ingested 3 capsules per day for 12 weeks. When recruiting subjects, the study was fully explained to the candidates, and informed consent was obtained. The candidates were also asked about their susceptibility to colds, and those who frequently caught colds were judged to be suitable subjects and were given priority for adoption. Stool samples were collected before and 12 weeks after the start of the study, and comprehensive analysis of the intestinal flora was performed in the same manner as in Example 1, and the occupancy rate of Agasobacter bacteria relative to the total number of eubacteria in the feces was measured.

[0033] 3. Test Results The occupancy rate of Agasobacter bacteria in the total number of eubacteria in the feces of the subjects before and after the start of the study is shown in Table 2. The unit is %.

[0034] [Table 2]

[0035] As shown in Table 2, it was confirmed that the intestinal occupancy rate of Agasobacter bacteria in the test diet group was significantly higher 12 weeks after the start of the study than before the start of the study. Therefore, it was found that the intake of capsule food containing Lactobacillus paragaceri bacteria promoted the increase of Agasobacter bacteria in healthy people who tend to catch colds easily.

[0036] (Examples of food additives) 100 mg of Lactobacillus paragaceri SBT2055 cells were mixed with 30 g of skim milk powder, 40 g of an equal mixture of vitamin C and citric acid, 100 g of granulated sugar, and 60 g of an equal mixture of cornstarch and lactose. The mixture was packed into a stick-shaped bag to produce the stick-shaped health food for promoting the growth of Agasobacter bacteria of the present invention.

[0037] (Example of mixing into feed) 20 g of Lactobacillus paragaceri SBT2055 cells were suspended in 3980 g of deionized water, heated to 40°C, and then stirred and mixed for 20 minutes at 3,600 rpm in a TK homomixer (MARK II 160 type; manufactured by Tokushu Kika Kogyo Co., Ltd.) to obtain a cell solution of 2 g / 4 kg. 2 kg of this cell solution was mixed with 1 kg of soybean meal, 1 kg of skimmed milk powder, 0.4 kg of soybean oil, 0.2 kg of corn oil, 2.3 kg of palm oil, 1 kg of corn starch, 0.9 kg of wheat flour, 0.2 kg of bran, 0.5 kg of vitamin mixture, 0.3 kg of cellulose, and 0.2 kg of mineral mixture, and sterilized at 120°C for 4 minutes to produce 10 kg of the feed for promoting the growth of Agasobacter bacteria of the present invention.

[0038] (Example of drug formulation) The liquid culture of Lactobacillus paragaceri SBT2055 was centrifuged at 7,000 rpm at 4°C for 15 minutes, and then washed with sterilized water and centrifuged three times to obtain washed cells. The washed cells were freeze-dried to obtain a cell powder. One part of the cell powder was mixed with four parts of skim milk powder, and the mixed powder was tableted in 1g portions using a tableting machine in a conventional manner to prepare the tablets for promoting the growth of Agasobacter bacteria of the present invention. [Industrial Applicability]

[0039] According to the present invention, it is possible to provide a composition for promoting the growth of Agasobacter bacteria, which contains as an active ingredient a bacterium and / or culture of Lactobacillus paragaceri, a food or beverage for promoting the growth of Agasobacter bacteria, a pharmaceutical for promoting the growth of Agasobacter bacteria, and a feed for promoting the growth of Agasobacter bacteria.

Claims

1. A composition for promoting the growth of bacteria of the genus Agasobacter, comprising as an active ingredient a bacterial cell and / or a culture product of Lactobacillus paragasseri.

2. The composition for promoting the growth of Agasobacter bacteria according to claim 1, wherein the Lactobacillus paragaceri is Lactobacillus paragaceri SBT2055 (FERM BP-10953).

3. 3. The composition for promoting the growth of Agasobacter bacteria according to claim 1 or 2, wherein the effect of promoting the growth of Agasobacter bacteria is to increase the occupancy rate of Agasobacter bacteria in human intestinal bacterial flora.

4. A food or drink for promoting the growth of Agasobacter bacteria, comprising the composition for promoting the growth of Agasobacter bacteria according to claim 1 or 2.

5. A pharmaceutical for promoting the growth of Agasobacter bacteria, comprising the composition for promoting the growth of Agasobacter bacteria according to claim 1 or 2.

6. A feed for promoting the growth of bacteria of the genus Agasobacter, comprising the composition for promoting the growth of bacteria of the genus Agasobacter according to claim 1 or 2.

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