Bifidobacteria growth promoter

Abstract

To provide an agent for increasing a ratio of bifidobacteria inside a human intestine, the bifidobacteria including Lactobacillus mucosae as active principle, and a bifidobacteria proliferation promotor, the bifidobacteria including Lactobacillus mucosae as active principle.SOLUTION: This invention relates to: an agent for increasing a ratio of bifidobacteria inside a human intestine, the bifidobacteria including Lactobacillus mucosae as an active principle; a bifidobacteria proliferation promotor, the bifidobacteria including Lactobacillus mucosae as an active principle; a beverage / food product for increasing a ratio of bifidobacteria inside the human intestine, the bifidobacteria including Lactobacillus mucosae as an active principle; and a beverage / food product for proliferating bifidobacteria including Lactobacillus mucosae as an active principle.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to a bifidobacteria growth promoter. In particular, the present invention relates to an agent for increasing the proportion of bifidobacteria in the human intestine and food and drink for promoting the increase, which contains Lactobacillus mucosae as an active ingredient. The present invention also relates to a bifidobacteria growth promoter and food and drink for promoting the growth, which contains Lactobacillus mucosae as an active ingredient. [Background technology]

[0002] There are 100 trillion bacteria in the human large intestine, forming the intestinal flora. The balance of bacterial species in this intestinal flora changes depending on health status and aging, and it has been reported that the proportion of bifidobacteria and lactic acid bacteria among intestinal bacteria decreases. One solution to supplement the decreasing bifidobacteria and lactic acid bacteria is to consume fermented dairy products that contain them. The representative types of lactic acid bacteria used in fermented dairy products are those belonging to the Lactobacillus genus. Commonly used species include Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus casei, Lactobacillus gasseri, Lactobacillus reuteri, and Lactobacillus helveticus. On the other hand, a species that is not often used is Lactobacillus mucosae. Patent Document 1 lists Lactobacillus mucosae as one of the lactic acid bacteria useful for improving lactose intolerance. The Lactobacillus mucosae in this patent document is characterized by its high intestinal adhesion and lactose decomposition ability, but does not disclose its effect on the ratio of bifidobacteria in the human large intestine or on the proliferation of bifidobacteria in a test tube. In addition, since lactose promotes the growth of bifidobacteria, it is possible that ingesting Lactobacillus mucosae, which has a high lactose decomposition ability, may reduce the proportion of bifidobacteria in the human large intestine or inhibit the growth of bifidobacteria. Non-patent documents 1 and 2 show that porcine-derived Lactobacillus mucosae LM1 has high intestinal adhesion and inhibits the adhesion of pathogenic bacteria to the intestinal tract. However, neither of these documents show the effect of Lactobacillus mucosae LM1 on bifidobacteria. Furthermore, Non-Patent Document 1 is an example of an in vitro analysis of adhesion to the intestinal tract, and does not show any effect in humans. Non-Patent Document 2 uses mice to analyze the inhibition of adhesion of pathogenic bacteria to the intestinal tract, and does not show any effect in humans. As described above, the effect of Lactobacillus mucosae in promoting the proliferation of bifidobacteria or increasing the proportion of bifidobacteria in the human large intestine has not been demonstrated, and the method by which it does so has been unknown. [Prior art documents] [Patent documents]

[0003] [Patent Document 1] International Publication No. WO2008 / 001676 [Non-patent literature]

[0004] [Non-Patent Document 1] J.Appl Microbiol. 2014 No. 117, Volume 2, pp. 485-497 [Non-Patent Document 2] Microb.Pathog.2019 No. 137 103760 Summary of the Invention [Problem to be solved by the invention]

[0005] The object of the present invention is to provide a new probiotic. In particular, the object is to provide an agent for increasing the ratio of bifidobacteria in the human intestine and a food and drink for increasing the ratio, which contains Lactobacillus mucosae as an active ingredient. The object of the present invention is also to provide an agent for promoting the growth of bifidobacteria and a food and drink for promoting the growth, which contains Lactobacillus mucosae as an active ingredient. [Means for solving the problem]

[0006] The present inventors have found for the first time that Lactobacillus mucosae has a growth promoting effect on bifidobacteria, and have confirmed that it can be used as a bifidobacteria growth promoter and a food and drink for promoting growth, and can be used as an agent for increasing the proportion of bifidobacteria in the human intestine and a food and drink for increasing the proportion of bifidobacteria in the human intestine, and have completed the present invention as a new probiotic. <1> An agent that increases the proportion of bifidobacteria in the human intestine, containing Lactobacillus mucosae as its active ingredient. <2> A bifidobacteria growth promoter containing Lactobacillus mucosae as the active ingredient. <3> When Lactobacillus mucosae is cultured with bifidobacteria, it has a proliferation activity 1.1 times higher than that of the untreated strain. <1> or <2> The agent described in. <4> Lactobacillus mucosae is one or more lactic acid bacteria strains selected from the group consisting of SBT2025 (NITE P-03189), SBT2268 (NITE P-03190), SBT2269 (NITE P-03191), SBT2867 (NITE P-03192), SBT10043 (NITE P-03187) and SBT10228 (NITE P-03188). <1> ~ <3> The agent according to any one of the preceding claims. <5> A food and beverage containing Lactobacillus mucosae as an active ingredient to increase the proportion of bifidobacteria in the human intestine. <6> A food and beverage that promotes the growth of bifidobacteria, containing Lactobacillus mucosae as an active ingredient. <7> When Lactobacillus mucosae is cultured with bifidobacteria, it has a proliferation activity 1.1 times higher than that of the untreated strain. <5> or <6> The food or beverage described in. <8> Lactobacillus mucosae is one or more lactic acid bacteria strains selected from the group consisting of SBT2025 (NITE P-03189), SBT2268 (NITE P-03190), SBT2269 (NITE P-03191), SBT2867 (NITE P-03192), SBT10043 (NITE P-03187) and SBT10228 (NITE P-03188). <5> ~ <7> 13. The food or beverage product according to any one of the preceding claims. <9> A method for producing a fermented dairy product, comprising a step of culturing Lactobacillus mucosae in a medium containing milk as a main component, characterized in that the Lactobacillus mucosae used is a strain having the following properties: Properties: Lactobacillus mucosae is characterized by having a proliferation activity that is 1.1 times higher when cultured with bifidobacteria than when no additives are added. <10> Lactobacillus mucosae is one or more lactic acid bacteria strains selected from the group consisting of SBT2025 (NITE P-03189), SBT2268 (NITE P-03190), SBT2269 (NITE P-03191), SBT2867 (NITE P-03192), SBT10043 (NITE P-03187) and SBT10228 (NITE P-03188). <9> The manufacturing method described in <11> A strain of Lactobacillus mucosae, comprising SBT2025 (NITE P-03189), SBT2269 (NITE P-03191), SBT2867 (NITE P-03192), SBT2268 (NITE P-03190), SBT10043 (NITE P-03187) or SBT10228 (NITE P-03188). Effect of the Invention

[0007] The present invention provides a fermented milk product containing Lactobacillus mucosae as an active ingredient, which has the activity of promoting the proliferation of bifidobacteria. By ingesting the fermented milk product containing Lactobacillus mucosae, the proportion of bifidobacteria in the human intestine can be increased. Therefore, the present invention makes it possible to provide a new probiotic. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0008] (Lactobacillus mucosae) In this specification, "Lactobacillus mucosae", which is an active ingredient of the agent for increasing the proportion of bifidobacteria in the human intestine and the agent for promoting the growth of bifidobacteria, means a bacterium belonging to Lactobacillus mucosae. That is, it refers to a strain whose 16S ribosomal RNA gene base sequence has a homology of 97% or more, more preferably 98% or more, and even more preferably 99% or more with the Lactobacillus mucosae type strain JCM12515. The source of isolation may be any, and more preferably it is derived from humans. The Lactobacillus mucosae strain used as an active ingredient in the present invention may be any strain that has the activity of promoting the growth of bifidobacteria, such as SBT2025, SBT2268, SBT2269, SBT2867, SBT10043, and SBT10228, whose activity is shown in the examples described below. Lactobacillus mucosae, which is the active ingredient of the agent for increasing the proportion of bifidobacteria and the agent for promoting the growth of bifidobacteria in this specification, may be in a state that has the activity of increasing the proportion of bifidobacteria or promoting the growth of bifidobacteria, and may be a live bacterial cell itself or a bacterial cell culture containing live bacterial cells. Examples of the live bacterial cell itself include a bacterial cell concentrate obtained by culturing and collecting the bacterial cells, and a dried product of the bacterial cells, and examples of the bacterial cell culture include fermented milk products of Lactobacillus mucosae, which will be described later. Lactobacillus mucosae can be cultured according to a conventional method. Various media can be used, such as a milk medium or a medium containing milk components, or a semi-synthetic medium that does not contain milk components. Examples of such media include reduced skim milk medium. The cells separated from the obtained culture by a collecting means such as centrifugation can be used as the active ingredient of the present invention as it is. Concentrated, dried, freeze-dried cells can also be used. Not only purely isolated cells, but also cultures, suspensions and other substances containing cells can be used. Examples of the form of the culture include, but are not limited to, cultures using media generally used for culturing lactic acid bacteria, as well as dairy products such as cheese, fermented milk, and dairy lactic acid bacteria drinks.

[0009] (Bifidobacteria) In the present specification, "bifidobacteria" refers to bacteria belonging to the genus Bifidobacterium. Bifidobacteria are not particularly limited as long as they are bacteria belonging to the genus Bifidobacterium, and examples thereof include Bifidobacterium longum, Bifidobacterium pseudocatenulatum, Bifidobacterium adolescentis, Bifidobacterium bifidum, Bifidobacterium breve, and Bifidobacterium dentium. Among these, a preferred example is Bifidobacterium longum. In addition, examples of the strain include Bifidobacterium longum type strain JCM1217, or Bifidobacterium dentium. An example of such a strain is Therium longum SBT2928 (accession number: FERM P-10657, date of deposit: April 13, 1989, National Institute of Advanced Industrial Science and Technology, Patent Organism Depositary Center).

[0010] (Fermented dairy products) In this specification, fermented milk products refer to those cultured in a liquid or paste containing milk or an equivalent or greater amount of non-fat milk solids, in which Lactobacillus mucosae has been cultivated. Milk may be raw milk, cow's milk, raw goat's milk, raw sheep's milk, etc., and skim milk powder may be reconstituted with water. When using skim milk powder, the concentration is not limited as long as the non-fat solids are equal to or greater than those of milk and the concentration is such that Lactobacillus mucosae can be cultivated, but it is desirable that the concentration be 8% (w / w) or more, preferably 9% (w / w) or more, and more preferably 10% (w / w) or more.

[0011] (Ingredients other than skimmed milk powder) The fermented milk product may contain nutrients other than skim milk powder, and when yeast extract is added, it is desirable that the amount is 0.1% (w / w) or more, preferably 0.2% (w / w) or more, more preferably 0.5% (w / w) or more. Other nutrients include sugars, amino acids, minerals, vitamins, etc. The fermented milk product of the present invention may contain bacteria other than Lactobacillus mucosae. For example, it may contain lactic acid bacteria commonly used as fermentation starters for fermented milk, such as Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus. The Lactobacillus mucosae of the present invention may also be provided as a mixed culture (e.g., fermented milk product) with bifidobacteria in which the proliferation is promoted by co-cultivating it with bifidobacteria.

[0012] (Lactobacillus mucosae viable cell count in fermented dairy products) It is desirable that Lactobacillus mucosae in fermented dairy products is contained as viable bacteria. In this specification, viable bacteria refers to colony formation in a solid medium containing nutrients on a support such as agar, and the number of viable bacteria refers to colony forming units (cfu) ( c olony f Orming u The viable cell count of Lactobacillus mucosae is expressed in units of nit. Examples of solid media for measuring the viable cell count of Lactobacillus mucosae include MRS agar medium for lactic acid bacteria and LBS agar medium for lactobacillus. The viable cell count of Lactobacillus mucosae in fermented dairy products is preferably 1.0E+06cfu / mL or more, more preferably 1.0E+07cfu / mL or more, and most preferably 1.0E+08cfu / mL or more.

[0013] (Promotes the growth of bifidobacteria) In this specification, "promoting the growth of bifidobacteria" refers to an increase in the number of live bifidobacteria. The activity of promoting the growth of bifidobacteria is said to be present when the increase in the number of live bifidobacteria is greater when Lactobacillus mucosae is added than when it is not added. For example, when the number of live bifidobacteria obtained when Lactobacillus mucosae is added is greater than the number of live bifidobacteria obtained when Lactobacillus mucosae is added in a test tube, the activity is said to be present. In the present invention, an agent having the activity is called a bifidobacteria growth promoter. The Lactobacillus mucosae used as the active ingredient in the bifidobacteria growth promoter of the present invention is preferably a strain that, when the viable cell count of bifidobacteria obtained when cultured without the addition of Lactobacillus mucosae is taken as 1.0 times, is preferably a strain that exhibits an increase in the viable cell count of bifidobacteria obtained when Lactobacillus mucosae is added by 1.1 times or more, more preferably 1.5 times or more, and even more preferably 2.0 times or more.

[0014] (Live count of bifidobacteria) The viable cell count of bifidobacteria refers to the number of colonies formed observed in GAM medium, which is an anaerobic bacterial medium in which bifidobacteria can grow, or TOS propionic acid agar medium, which is a medium for bifidobacteria, etc.

[0015] (Increasing the proportion of bifidobacteria in the human intestine) In this specification, "increasing the proportion of bifidobacteria in the human intestine" refers to increasing the proportion of bifidobacteria in the human intestinal bacteria. For example, when the proportion of bifidobacteria in the intestinal bacteria after ingestion of Lactobacillus mucosae is greater than 1.0 times that before ingestion, it is said to be active. In the present invention, an agent having this activity is called an agent for increasing the proportion of bifidobacteria in the intestine. The Lactobacillus mucosae that is the active ingredient of the agent for increasing the proportion of bifidobacteria in the intestine of the present invention is preferably a strain that shows a proportion of 1.1 times or more after ingestion compared to before ingestion, more preferably 1.3 times or more, even more preferably 2.0 times or more, and even more preferably 3.0 times or more. One method for screening such strains is to first screen for strains that show a 1.1-fold or higher viable cell count when Lactobacillus mucosae is added, when compared to 1.0-fold viable cell count when bifidobacteria are cultured in a test tube under culture conditions that mimic the intestinal environment, without the addition of Lactobacillus mucosae.

[0016] (Increased proportion of bifidobacteria) The percentage of bifidobacteria in the human intestine means the percentage of bifidobacteria in the DNA in feces. The 16S rRNA gene of DNA obtained from feces can be amplified by PCR, and the percentage of DNA derived from bifidobacteria in the entire PCR product can be calculated. The percentage can be calculated using meta-16S analysis with a next-generation sequencer or T-RFLP ( T terminal R striction F ragment L ength P Microbial flora analysis techniques such as the olymorphism method can be used.

[0017] (Agent for increasing the proportion of bifidobacteria in the human intestine and for promoting the proliferation of bifidobacteria) The agent for increasing the proportion of bifidobacteria in the human intestine and the agent for promoting the proliferation of bifidobacteria of the present invention can utilize the fermentation culture of Lactobacillus mucosae itself or the bacteria itself. Examples of the fermentation culture used as a food include the above-mentioned fermented dairy products. Furthermore, it can be used after being formulated. When preparing a formulation, it can be mixed with excipients, stabilizers, flavorings, etc. permitted for pharmaceutical use, concentrated, freeze-dried, or heated and dried to kill the bacteria. These dried products, concentrates, and paste-like products are also included. In addition, the composition may be formulated by mixing excipients, binders, disintegrants, lubricants, flavorings, suspending agents, coating agents, and other optional agents within the range that does not impair the bifidobacterium growth-promoting activity. The dosage form may be tablets, capsules, granules, powders, dust, syrup, etc., and these are preferably administered orally.

[0018] (Food and drink for increasing the proportion of bifidobacteria in the human intestine and food and drink for promoting the proliferation of bifidobacteria) The food and drink for increasing the proportion of bifidobacteria in the human intestine and the food and drink for promoting the proliferation of bifidobacteria of the present invention can utilize the fermentation culture of Lactobacillus mucosae itself. Examples of the fermentation culture used as a food include the above-mentioned fermented dairy products. In addition, these foods and beverages may be prepared by blending the agent for increasing the proportion of bifidobacteria in the human intestine and the agent for promoting the proliferation of bifidobacteria into suitable foods and beverages. These may be added to the raw materials during the manufacturing process of the foods and beverages, may be blended into the ingredients, or may be blended into the final foods and beverages. Examples of foods and beverages include, but are not limited to, dairy products such as cheese, fermented milk, dairy lactic acid bacteria drinks, lactic acid bacteria drinks, butter, margarine, and the like, beverages such as milk drinks, fruit juice drinks, and soft drinks, egg products such as jelly, candy, pudding, and mayonnaise, confectioneries and breads such as butter cake, and further various types of milk powder, as well as foods for infants and young children, and nutritional compositions. Furthermore, the foods and beverages of the present invention for increasing the proportion of bifidobacteria in the human intestine and for promoting the proliferation of bifidobacteria can also be used as functional foods, foods for specified health uses, foods with nutrient function claims, and foods for beauty.

[0019] When preparing an agent for increasing the proportion of bifidobacteria in the human intestine, a bifidobacteria growth promoter, a food or drink for increasing the proportion of bifidobacteria in the human intestine, a food or drink for promoting the growth of bifidobacteria, a nutritional composition for increasing the proportion of bifidobacteria in the human intestine, or a nutritional composition for promoting the growth of bifidobacteria, by adding Lactobacillus mucosae bacteria and / or culture, the mixing ratio is not particularly limited and may be appropriately adjusted according to the ease of production and the preferred daily dosage. It is determined individually taking into consideration the symptoms and age of the person to be administered, but the mixing amount may be adjusted so that an adult can usually ingest 10 to 200 g of Lactobacillus mucosae bacteria culture or 0.1 to 100 mg of the bacteria itself. Examples of the present invention will be described in detail below, but the present invention is not limited to these examples. EXAMPLES

[0020] [Example 1] Promoting the growth of bifidobacteria (1) Strains For Lactobacillus mucosae, we used the strains isolated from humans, as shown in Table 1. For bifidobacteria, we used the type strain JCM1217, which is the same species as Bifidobacterium longum, known to be widely distributed in Japanese people from infants to the elderly.

[0021] [Table 1]

[0022] (2) Preparation of concentrated Lactobacillus mucosae cells MRS liquid medium for lactic acid bacteria was prepared and sterilized by heating at 121°C for 15 minutes. Lactobacillus mucosae SBT2025, Lactobacillus mucosae SBT2268, Lactobacillus mucosae SBT2269, Lactobacillus mucosae SBT2867, Lactobacillus mucosae SBT10043, and Lactobacillus mucosae SBT10228 shown in Table 1 were inoculated into the medium and cultured anaerobically at 37°C for 16 hours using an anaerobic culture system (product name: Anaeropack, Mitsubishi Gas Chemical Co., Ltd.). The culture was concentrated by centrifugation and glycerol was added to a concentration of 10% (v / v). These concentrated bacterial cells were frozen at -80°C and then thawed, and serially diluted. They were then spread on MRS agar medium to measure the viable bacterial count, and it was confirmed that each strain contained a viable bacterial count of 2.0E+09 cfu / mL or more. The above strains were deposited at the Patent Microorganisms Depositary Center of the National Institute of Technology and Evaluation (2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture (postal code 292-0818)) on March 27, 2020. The deposit numbers are as follows: SBT2025: Accession number NITE P-03189 SBT2268: Accession number NITE P-03190 SBT2269: Accession number NITE P-03191 SBT2867: Accession number NITE P-03192 SBT10043: Accession number NITE P-03187 SBT10228: Accession number NITE P-03188

[0023] (3) Preparation of Lactobacillus mucosae-fermented dairy products A 10% (w / w) skim milk powder containing 0.5% (w / w) yeast extract and 2.5% (w / w) glucose was prepared and sterilized by heat treatment at 115°C for 20 minutes. The concentrated bacterial cells prepared as described above were added to the milk powder at 0.1% (v / w) and cultured anaerobically at 37°C for 16 hours using an anaerobic culture system. The resulting fermented milk product was serially diluted in volume and smeared on LBS agar medium for lactobacilli containing 0.132% (v / v) acetic acid to measure the viable bacterial count. It was confirmed that each strain contained a viable bacterial count of 1.0E+06 cfu / mL or more. These were compared with Bifidobacterium longum JCM1217. Used in mixed cultures.

[0024] (4) Preparation of concentrated Bifidobacterium longum JCM1217 cells GAM liquid medium (GAM bouillon, product code 05422, manufactured by Nissui Pharmaceutical Co., Ltd.), a medium for anaerobic bacteria, was prepared and sterilized by heating at 115°C for 15 minutes. Bifidobacterium longum JCM1217 was inoculated into the medium and anaerobically cultured at 37°C for 16 hours using an anaerobic culture system. The resulting culture was concentrated by centrifugation, and glycerol was added to a concentration of 10% (v / v) to obtain concentrated bacterial cells. These concentrated bacterial cells were frozen at -80°C and then thawed, after which they were serially diluted and smeared on TOS propionic acid agar medium for bifidobacteria to measure the viable bacterial count, which was 1.0E+09 cfu / mL. This was used in a mixed culture with Lactobacillus mucosae.

[0025] (5) Mixed culture of Lactobacillus mucosae fermented dairy product and Bifidobacterium longum JCM1217 GAM liquid medium (GAM bouillon, product code 05422, manufactured by Nissui Pharmaceutical Co., Ltd.), a medium for anaerobic bacteria, was prepared and sterilized by heating at 115°C for 15 minutes. 1% (v / v) Lactobacillus mucosae fermented dairy product (1.0E+06cfu / mL or more) and 10% (v / v) Bifidobacterium longum JCM1217 concentrated bacteria (1.0E+09cfu / mL) were added and mixed, and cultured at 37°C for 16 hours using an anaerobic culture system. As a control, a test group was set up where no Lactobacillus mucosae fermented dairy product was added. GAM liquid medium is used as a model medium for the intestinal environment. It is a culture medium (PLOS ONE DOI:10.1371 / JOURNAL.PONE.0160533 August2,2016).

[0026] (6) Bifidobacterial count and proliferation after mixed culture The mixed culture was serially diluted 10-fold in volume, spread on a TOS propionic acid agar medium containing 0.005% (w / v) mupirocin lithium, which is a selective medium for bifidobacteria, and cultured for 3 days at 37°C using an anaerobic culture system. The number of colonies formed on the agar medium was observed, and the increase in the number of colonies formed (cfu) by each fermented dairy product, i.e., the promotion of bifidobacterial growth, was calculated as a factor, relative to the control group, which was set at 1.0.

[0027] As a result, the number of viable bifidobacteria in the control group without addition of Lactobacillus mucosae fermented milk product was 4.7E+08cfu / mL, whereas the number was 8.7E+08cfu / mL (1.8-fold increase in growth) in the group with addition of SBT2025 fermented milk product, 1.0E+09cfu / mL (2.1-fold increase in growth) in the group with addition of SBT2268 fermented milk product, 1.0E+09cfu / mL (2.1-fold increase in growth) in the group with addition of SBT2269 fermented milk product, 1.3E+09cfu / mL (2.9-fold increase in growth) in the group with addition of SBT2867 fermented milk product, 5.1E+08cfu / mL (1.1-fold increase in growth) in the group with addition of SBT10043 fermented milk product, and 1.3E+09cfu / mL (2.7-fold increase in growth) in the group with addition of SBT10228 fermented milk product. In other words, these Lactobacillus mucosae fermented dairy products promoted the proliferation of bifidobacteria by 1.1 times or more compared to the control group (Table 2). 8 " " x 10 9 ".

[0028] [Table 2]

[0029] [Example 2] Increasing the proportion of bifidobacteria in the human intestine (1) Bacterial strains and fermented dairy product trials For SBT10043, which showed a 1.1-fold increase in growth in Example 1, an edible fermented milk product was produced. After isolating SBT10043 as a single colony on an MRS agar medium, the bacteria were picked up in a nutrient medium consisting of 1% (w / w) edible yeast extract, 2% (w / w) edible casein peptone, and 2% (w / w) edible glucose, and anaerobically cultured at 37°C for 16 hours using an anaerobic culture system. This culture was concentrated 10 times in volume by centrifugation to obtain edible concentrated bacterial cells. Next, 10% (w / w) skim milk powder containing 0.5% (w / w) edible yeast extract and 2.5% (w / w) edible glucose was prepared and sterilized at 115°C for 20 minutes. 10% (v The edible concentrated cells were added so that the total volume was 100 ml / w, and the mixture was incubated at 37°C for 24 hours in a sealed container.

[0030] The above fermented milk product was inoculated to 10% (w / w) skim milk powder containing 0.5% (w / w) edible yeast extract and 2.5% (w / w) edible glucose, which had been freshly prepared and sterilized, at 10% (w / w), and the mixture was allowed to stand at 37°C for 24 hours in a sealed container. This was used as an edible fermented milk product. The edible fermented milk product was also produced multiple times as necessary by repeating subculture using the same edible fermented milk product as a seed culture. That is, the edible fermented milk product was inoculated to 10% (w / w) in the same skim milk medium (10% (w / w) skim milk powder containing 0.5% (w / w) edible yeast extract and 2.5% (w / w) edible glucose) and the mixture was allowed to stand at 37°C for 24 hours in a sealed container. The strain was repeatedly subcultured by static cultivation for 24 hours to be used as seed culture for edible fermented milk products, and the culture volume was increased as necessary to be used for consumption as edible fermented milk products.

[0031] (2) Viable cell count of SBT10043 in edible fermented dairy products 1.0 mL of the edible fermented dairy product was serially diluted to 10 times the volume, and smeared on LBS agar medium for lactobacilli containing 0.132% (v / v) acetic acid to measure the viable bacterial count, confirming that it contained a viable bacterial count of 5.0E+08 cfu / mL or more.

[0032] (3) Duration and consumers of edible fermented dairy products The edible fermented dairy product containing the above SBT10043 was consumed at 200 mL per day, i.e., at least 1.0E+09 cfu / day, for 14 days. Feces were collected before and after consumption, DNA was extracted, and the proportion of bifidobacteria was determined using the T-RFLP method, a bacterial flora analysis method. The subjects were two people: Subject A, who had less than 10% bifidobacteria before consuming the edible fermented dairy product, and Subject B, who had more than 10% bifidobacteria before consuming the edible fermented dairy product.

[0033] (4) Increase in the proportion of intestinal bifidobacteria in consumer A The percentage of bifidobacteria in the large intestine of consumer A before ingesting the edible fermented dairy product containing Lactobacillus mucosae was 3.3%, but after ingesting the fermented dairy product for 14 days, it increased to 11.6%, i.e., 3.5-fold increase after ingestion of the fermented dairy product compared to before ingestion (Table 3).

[0034] [Table 3]

[0035] (5) Increase in the proportion of intestinal bifidobacteria in consumer B The percentage of bifidobacteria in the large intestine of consumer B before ingesting the edible fermented dairy product containing Lactobacillus mucosae was 19.9%, but after ingesting the fermented dairy product for 14 days, it increased to 25.9%, i.e., 1.3-fold increase after ingestion of the fermented dairy product compared to before ingestion (Table 4). From the above, it was found in Test Example 1 that the proportion of bifidobacteria in the large intestine can be increased by humans ingesting fermented dairy products containing Lactobacillus mucosae, which showed growth-promoting activity for bifidobacteria in a culture test that mimicked the human large intestine environment.

[0036] [Table 4] [Industrial Applicability]

[0037] According to the present invention, the proportion of bifidobacteria in the human large intestine can be increased by ingesting a fermented milk product of Lactobacillus mucosae, which promotes the proliferation of bifidobacteria in vitro, making it possible to provide a new probiotic.

Claims

1. An agent for increasing the proportion of bifidobacteria in the human intestine, containing Lactobacillus mucosae as an active ingredient (excluding the above-mentioned agents that contain all of Lactobacillus plantarum, Faecalibacterium prausnitzii, Butyricicoccus pullicaecorum, Roseburia inulinivorans, Roseburia hominis, Akkermansia muciniphila, Anaerostipes caccae, Escherichia coli, Enterococcus faecium, Lactobacillus mucosae, Bifidobacterium adolescentis, Bifidobacterium longum, Bacteroides thetaiotaomicron, and Bacteroides vulgatus).

2. A bifidobacterial growth promoter containing Lactobacillus mucosae as the active ingredient (excluding the above-mentioned agents that contain all of Lactobacillus plantarum, Faecalibacterium prausnitzii, Butyricicoccus pullicaecorum, Roseburia inulinivorans, Roseburia hominis, Akkermansia muciniphila, Anaerostipes caccae, Escherichia coli, Enterococcus faecium, Lactobacillus mucosae, Bifidobacterium adolescentis, Bifidobacterium longum, Bacteroides thetaiotaomicron, and Bacteroides vulgatus).

3. 3. The agent according to claim 1 or 2, wherein Lactobacillus mucosae is a strain that has a proliferation activity that is 1.1 times or more higher when cultured with bifidobacteria than when no agent is added.

4. The agent according to any one of claims 1 to 3, wherein Lactobacillus mucosae is one or more lactic acid bacteria strains selected from the group consisting of SBT2025 (NITE P-03189), SBT2268 (NITE P-03190), SBT2269 (NITE P-03191), SBT2867 (NITE P-03192), SBT10043 (NITE P-03187) and SBT10228 (NITE P-03188).

5. Food and beverage products for increasing the proportion of bifidobacteria in the human intestine, containing Lactobacillus mucosae as an active ingredient (excluding the above foods and beverage products that contain all of Lactobacillus plantarum, Faecalibacterium prausnitzii, Butyricicoccus pullicaecorum, Roseburia inulinivorans, Roseburia hominis, Akkermansia muciniphila, Anaerostipes caccae, Escherichia coli, Enterococcus faecium, Lactobacillus mucosae, Bifidobacterium adolescentis, Bifidobacterium longum, Bacteroides thetaiotaomicron, and Bacteroides vulgatus).

6. Food and beverage products for promoting the growth of bifidobacteria that contain Lactobacillus mucosae as an active ingredient (however, excluding the above foods and beverage products that contain all of Lactobacillus plantarum, Faecalibacterium prausnitzii, Butyricicoccus pullicaecorum, Roseburia inulinivorans, Roseburia hominis, Akkermansia muciniphila, Anaerostipes caccae, Escherichia coli, Enterococcus faecium, Lactobacillus mucosae, Bifidobacterium adolescentis, Bifidobacterium longum, Bacteroides thetaiotaomicron, and Bacteroides vulgatus).

7. 7. The food or beverage according to claim 5 or 6, wherein Lactobacillus mucosae is a strain that has a proliferation activity that is 1.1 times or more higher when cultured with bifidobacteria than when not added.

8. The food or drink according to any one of claims 5 to 7, wherein the Lactobacillus mucosae is one or more lactic acid bacteria strains selected from the group consisting of SBT2025 (NITE P-03189), SBT2268 (NITE P-03190), SBT2269 (NITE P-03191), SBT2867 (NITE P-03192), SBT10043 (NITE P-03187) and SBT10228 (NITE P-03188).

9. A method for producing a fermented dairy product (excluding the fermented dairy product containing all of Lactobacillus plantarum, Faecalibacterium prausnitzii, Butyricicoccus pullicaecorum, Roseburia inulinivorans, Roseburia hominis, Akkermansia muciniphila, Anaerostipes caccae, Escherichia coli, Enterococcus faecium, Lactobacillus mucosae, Bifidobacterium adolescentis, Bifidobacterium longum, Bacteroides thetaiotaomicron and Bacteroides vulgatus), which comprises a step of culturing Lactobacillus mucosae in a medium containing milk as the main component, wherein the method is characterized in that a strain of Lactobacillus mucosae having the following properties is used: Properties: Lactobacillus mucosae is characterized by having a proliferation activity 1.1 times or more higher than that of the control when cultured with bifidobacteria.

10. The method according to claim 9, wherein the Lactobacillus mucosae is one or more lactic acid bacteria strains selected from the group consisting of SBT2025 (NITE P-03189), SBT2268 (NITE P-03190), SBT2269 (NITE P-03191), SBT2867 (NITE P-03192), SBT10043 (NITE P-03187) and SBT10228 (NITE P-03188).

11. A strain of Lactobacillus mucosae, which is SBT2025 (NITE P-03189), SBT2269 (NITE P-03191), SBT2867 (NITE P-03192), SBT2268 (NITE P-03190), SBT10043 (NITE P-03187) or SBT10228 (NITE P-03188).

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