Composition for promoting secretion of galactotropin

Abstract

The present invention addresses the problem of providing a novel composition for promoting galactotropin secretion. A solution is provided. Provided are: a composition for promoting the secretion of gastro-somatotropin, which contains a bacterium belonging to Lactobacillus delbrueckii; and a composition for promoting the secretion of gastro-somatotropin, which is used for promoting the secretion of growth hormone, inhibiting the function of brown adipose tissue, promoting the movement of the digestive tract, increasing the secretion of gastric acid, increasing the secretion of pancreas exocrine, and expanding blood vessels. The composition is one or more of the group consisting of a composition for increasing cardiac contractility and protecting cardiac muscle cells.

Description

Technical Field

[0001] The present invention relates to a composition for promoting ghrelin secretion. Background Art

[0002] Ghrelin is a peptide hormone mainly secreted by gastric endocrine cells, and various functions have been reported, such as regulating energy metabolism, promoting gastric acid secretion and gastrointestinal motility, dilating blood vessels, protecting the cardiovascular system, and having anti-diabetic effects.

[0003] Somatolin has the effect of promoting the secretion of growth hormone. Growth hormone and somatolin are peptide hormones. When directly taken orally, they are decomposed by digestive enzymes and cannot exert their effects. In addition, injection of growth hormone has various problems such as stimulant problems, safety, and side effects. Therefore, a method that is highly safe, can be easily taken, and can promote the secretion of endogenous growth hormone is desired.

[0004] Patent documents 1 and 2 record hormone secretion promoters with skim milk or skim milk powder as active ingredients, and the hormones mentioned in these are recorded as ghrelin or growth hormone. In addition, patent document 3 records a ghrelin secretion promoter, which contains whey protein hydrolyzate and fermented milk protein as protein, medium-chain fatty acids, milk lecithin and fish oil as lipids, and palatinose (registered trademark) as sugar, and patent document 4 records an appetite improver, which contains Lactobacillus gasseri OLL2716 (FERM BP-6999) (reclassified as Lactobacillus paragasseri in 2020.) As an active ingredient, and records that the appetite improvement is based on the ghrelin secretion promoting effect. In addition, patent document 5 records peptides with specific sequences of 4 to 12 amino acids in length derived from subtilisin digests of purified rice endosperm protein, and records that these peptides are effective for treating anorexia caused by decreased ghrelin secretion.

[0005] On the other hand, Lactobacillus delbrueckii is a kind of lactic acid bacteria. Patent Document 6 records a lipid metabolism improver, which is a lipid metabolism improver containing a bacterial cell destruction product of lactic acid bacteria as an effective ingredient, wherein the average long diameter of the bacterial cell in the bacterial cell destruction product is 0 to 70% of that before destruction, and Lactobacillus delbrueckii subspecies bulgaricus is cited as an example of lactic acid bacteria. Patent Document 7 records a fermented milk for improving fatigue prepared using lactobacillus lactic acid bacteria, and as a lactic acid bacteria of the genus Lactobacillus, Lactobacillus delbrueckii subspecies bulgaricus OLL1073R-1 is recorded. Patent Document 8 describes a fermented milk for improving autonomic nervous function, which contains at least one lactic acid bacterium of the genus Lactobacillus selected from the group consisting of Lactobacillus delbrueckii, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus gasseri, Lactobacillus rhamnosus and Lactobacillus acidophilus, and at least one of their metabolites, wherein Lactobacillus delbrueckii subsp. bulgaricus OLL1073R-1 (FERM BP-10741) is described as the lactic acid bacterium of the genus Lactobacillus. Patent Document 9 discloses that a probiotic compound VSL is administered to improve autonomic nervous function. # 3. It was observed that the weight loss of the ulcerative colitis mouse model was inhibited, the disease activity index (DAI) was reduced, and the intestinal lesions were improved. The probiotic compound VSL # 3 contains 8 useful bacteria including Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus casei, Lactobacillus plantarum, Lactobacillus acidophilus, Streptococcus thermophilus, Bifidobacterium longum, Bifidobacterium breve and Bifidobacterium infantis. Non-patent document 1 records that Lactobacillus delbrueckii LAB4 inhibits the excessive response of insulin supplementation secretion caused by the early rise in blood sugar level just after sugar intake, thereby inhibiting hypoglycemia.

[0006] Prior art literature

[0007] Patent Literature

[0008] Patent Document 1: Japanese Patent Application Laid-Open No. 2015-205829

[0009] Patent Document 2: Japanese Patent Application Laid-Open No. 2015-205830

[0010] Patent Document 3: Japanese Patent No. 6657084 (International Publication No. WO2015 / 199192)

[0011] Patent Document 4: Japanese Patent No. 6813974 (Japanese Patent Application Laid-Open No. 2018-8901)

[0012] Patent Document 5: International Publication No. WO2020 / 218450

[0013] Patent Document 6: Japanese Patent No. 5801802 (International Publication No. WO2011 / 063117)

[0014] Patent Document 7: International Publication WO2017 / 015407

[0015] Patent Document 8: Japanese Unexamined Patent Application Publication No. 2020-184906

[0016] Patent Document 9: Japanese Patent Application Laid-Open for International Application No. 2021-524751

[0017] Non-Patent Document

[0018] Non-Patent Document 1: Shoutaro Nagata, Chika Otsuki, Satoako Shika, Mamoru Maekawa, Tetsuya Sugaya, Yurie Ikeuchi, Hypothesis on the mechanism of action of lactic acid bacteria LAB4 and lactic acid bacteria LAB4 supplemented with glucomannan against blood glucose spikes, Comprehensive Medicine 20(1)3-12, 2021. Summary of the Invention

[0019] Problems to be Solved by the Invention

[0020] The object of the present invention is to provide a novel composition for promoting the secretion of ghrelin.

[0021] Solutions to the Problems

[0022] The present invention relates to the following.

[0023] <1> Use of a bacterium belonging to Lactobacillus delbrueckii in the manufacture of a composition for promoting the secretion of ghrelin for therapeutic or non-therapeutic use.

[0024] <2> Use of a bacterium belonging to Lactobacillus delbrueckii in the manufacture of a composition for any one or more selected from the group consisting of promoting the secretion of growth hormone, inhibiting the function of brown adipose tissue, promoting gastrointestinal motility, enhancing gastric acid secretion, enhancing pancreatic exocrine secretion, dilating blood vessels, increasing cardiac contractility, and protecting cardiomyocytes.

[0025] <3> The use according to <2>, wherein any one or more selected from the group consisting of promoting the secretion of growth hormone, inhibiting the function of brown adipose tissue, promoting gastrointestinal motility, enhancing gastric acid secretion, enhancing pancreatic exocrine secretion, dilating blood vessels, increasing cardiac contractility, and protecting cardiomyocytes is based on promoting the secretion of ghrelin.

[0026] <4> The use according to any one of <1> to <3>, wherein the bacterium belonging to Lactobacillus delbrueckii is a bacterium belonging to any one of Lactobacillus delbrueckii ssp.bulgaricus, Lactobacillus delbrueckii ssp.delbrueckii, Lactobacillus delbrueckii ssp.indicus, Lactobacillus delbrueckii ssp.lactis and Lactobacillus delbrueckii ssp.sunkii.

[0027] <5> The use according to any one of <1> to <4>, wherein the bacterium belonging to Lactobacillus delbrueckii has a 16S rRNA gene having 90% or more identity with the base sequence represented by any one of SEQ ID NOs: 1 to 5.

[0028] <6> The use according to any one of <1> to <5>, wherein the bacteria belonging to Lactobacillus delbrueckii subsp. bulgaricus is selected from Lactobacillus delbrueckii subsp. bulgaricus 2038, Lactobacillus delbrueckii subsp. bulgaricus OLL1255 (Accession No.: NITE BP-76), Lactobacillus delbrueckii subsp. bulgaricus OLL1171 (Accession No.: NITE BP-01569), Lactobacillus delbrueckii subsp. bulgaricus OLL1247 (Accession No.: NITE BP-01814), Lactobacillus delbrueckii subsp. bulgaricus OLL205013 (Accession No.: NITE BP-02411), Lactobacillus delbrueckii OLL204989 (Accession No.: NITE BP-02874), Lactobacillus delbrueckii subsp. delbrueckii (JCM 1012T), Lactobacillus delbrueckii subsp. indica (JCM 15610T), Lactobacillus delbrueckii subsp. lactis (JCM Any one or more of the group consisting of Lactobacillus delbrueckii subsp. mori (JCM 17838T).

[0029] <7> The use according to any one of <1> to <6>, wherein the bacteria belonging to Lactobacillus delbrueckii are living bacteria.

[0030] <8> The use according to any one of <1> to <7>, wherein the composition is fermented milk that does not contain bacteria belonging to Lactobacillus paragaseri.

[0031] <9> The use according to any one of <1> to <8>, wherein the bacterium belonging to Lactobacillus delbrueckii does not have a DNA consisting of the sequence of SEQ ID NO: 6.

[0032] <10> Non-therapeutic use of a composition containing bacteria belonging to Lactobacillus delbrueckii for promoting ghrelin secretion.

[0033] <11> A composition containing bacteria belonging to Lactobacillus delbrueckii, for non-therapeutic use of any one or more selected from the group consisting of promoting growth hormone secretion, inhibiting the function of brown adipose tissue, promoting gastrointestinal motility, increasing gastric acid secretion, increasing pancreatic exocrine secretion, dilating blood vessels, increasing cardiac contractility and protecting cardiomyocytes.

[0034] <12> The use according to <11>, wherein the method comprises promoting the secretion of growth hormone, inhibiting the function of brown adipose tissue, promoting gastrointestinal motility, increasing gastric acid secretion, increasing pancreatic exocrine secretion, dilating blood vessels, increasing cardiac contractility and protecting cardiomyocytes, and any one or more thereof is based on promoting the secretion of ghrelin.

[0035] <13> The use according to any one of <10> to <12>, wherein the bacterium belonging to Lactobacillus delbrueckii is a bacterium belonging to any one of Lactobacillus delbrueckii ssp.bulgaricus, Lactobacillus delbrueckii ssp.delbrueckii, Lactobacillus delbrueckii ssp.indicus, Lactobacillus delbrueckii ssp.lactis and Lactobacillus delbrueckii ssp.sunkii.

[0036] <14> The use according to any one of <10> to <13>, wherein the bacterium belonging to Lactobacillus delbrueckii has a 16S rRNA gene that is 90% or more identical to the base sequence represented by any one of SEQ ID NOs: 1 to 5.

[0037] <15> The use according to any one of <10> to <14>, wherein the bacteria belonging to Lactobacillus delbrueckii subsp. bulgaricus is selected from Lactobacillus delbrueckii subsp. bulgaricus 2038, Lactobacillus delbrueckii subsp. bulgaricus OLL1255 (Accession No.: NITE BP-76), Lactobacillus delbrueckii subsp. bulgaricus OLL1171 (Accession No.: NITE BP-01569), Lactobacillus delbrueckii subsp. bulgaricus OLL1247 (Accession No.: NITE BP-01814), Lactobacillus delbrueckii subsp. bulgaricus OLL205013 (Accession No.: NITE BP-02411), Lactobacillus delbrueckii OLL204989 (Accession No.: NITE BP-02874), Lactobacillus delbrueckii subsp. delbrueckii (JCM 1012T), Lactobacillus delbrueckii subsp. indica (JCM 15610T), Lactobacillus delbrueckii subsp. lactis (JCM Any one or more of the group consisting of Lactobacillus delbrueckii subsp. mori (JCM 17838T).

[0038] <16> The use according to any one of <10> to <15>, wherein the bacteria belonging to Lactobacillus delbrueckii are living bacteria.

[0039] <17> The use according to any one of <10> to <17>, wherein the composition contains bacteria belonging to Lactobacillus delbrueckii.

[0040] <18> The use according to any one of <10> to <17>, wherein the composition is fermented milk that does not contain bacteria belonging to Lactobacillus paragaseri.

[0041] <19> The use according to any one of <10> to <18>, wherein the bacterium belonging to Lactobacillus delbrueckii does not have a DNA consisting of the sequence of SEQ ID NO: 6.

[0042] In addition, the present invention relates to the following.

[0043] [1] A composition for promoting ghrelin secretion, comprising bacteria belonging to the group consisting of Lactobacillus delbrueckii.

[0044] [2] A composition for use in any of the following: promoting growth hormone secretion, promoting fat accumulation (inhibiting weight loss), inhibiting insulin secretion, inhibiting the function of brown adipose tissue, promoting gastrointestinal motility, promoting gastric acid secretion, promoting pancreatic exocrine secretion, dilating blood vessels, increasing cardiac contractility, and protecting myocardial cells, comprising a bacterium belonging to Lactobacillus delbrueckii.

[0045] [3] The composition according to [2], which is based on promoting ghrelin secretion.

[0046] [4] The composition according to any one of [1] to [3], wherein the bacterium belonging to Lactobacillus delbrueckii is a bacterium belonging to any one of Lactobacillus delbrueckii ssp.bulgaricus, Lactobacillus delbrueckii ssp.delbrueckii, Lactobacillus delbrueckii ssp.indicus, Lactobacillus delbrueckii ssp.lactis and Lactobacillus delbrueckii ssp.sunkii.

[0047] [5] The composition according to any one of [1] to [4], wherein the bacteria belonging to Lactobacillus delbrueckii subsp. bulgaricus is selected from Lactobacillus delbrueckii subsp. bulgaricus 2038, Lactobacillus delbrueckii subsp. bulgaricus OLL1255 (Accession No.: NITE BP-76), Lactobacillus delbrueckii subsp. bulgaricus OLL1171 (Accession No.: NITE BP-01569), Lactobacillus delbrueckii subsp. bulgaricus OLL1247 (Accession No.: NITE BP-01814), Lactobacillus delbrueckii subsp. bulgaricus OLL205013 (Accession No.: NITE BP-02411), Lactobacillus delbrueckii OLL204989 (Accession No.: NITE BP-02874), Lactobacillus delbrueckii subsp. delbrueckii (JCM 1012T), Lactobacillus delbrueckii subsp. indica (JCM 15610T), Lactobacillus delbrueckii subsp. lactis (JCM Any one or more of the group consisting of Lactobacillus delbrueckii subsp. mori (JCM 17838T).

[0048] [6] The composition according to any one of [1] to [5], wherein the bacteria belonging to Lactobacillus delbrueckii are living bacteria.

[0049] [7] The composition according to any one of [1] to [6], which is fermented milk that does not contain bacteria belonging to Lactobacillus paragaseri.

[0050] [8] The composition according to any one of [1] to [7], wherein the bacteria belonging to Lactobacillus delbrueckii does not have a DNA consisting of the sequence of SEQ ID NO: 6.

[0051] In addition, the present invention relates to the following.

[0052] [9] A composition containing bacteria belonging to Lactobacillus delbrueckii, and a method for promoting the secretion of ghrelin. Use of bacteria belonging to Lactobacillus delbrueckii in the manufacture of a composition for promoting the secretion of ghrelin. A method or non-therapeutic method for promoting the secretion of ghrelin, comprising the step of administering a composition containing bacteria belonging to Lactobacillus delbrueckii to a subject. Use or non-therapeutic use of a composition containing bacteria belonging to Lactobacillus delbrueckii for promoting the secretion of ghrelin.

[0053]

[10] A composition containing bacteria belonging to Lactobacillus delbrueckii, which is used for any method selected from the group consisting of promoting growth hormone secretion, promoting fat accumulation (inhibiting weight loss), inhibiting insulin secretion, inhibiting the function of brown adipose tissue, promoting digestive tract motility, causing hypersecretion of gastric acid, causing hypersecretion of pancreatic exocrine secretion, dilating blood vessels, increasing cardiac contractility, and protecting cardiomyocytes. Use of bacteria belonging to Lactobacillus delbrueckii in the manufacture of a composition for any method selected from the group consisting of promoting growth hormone secretion, promoting fat accumulation (inhibiting weight loss), inhibiting insulin secretion, inhibiting the function of brown adipose tissue, promoting digestive tract motility, causing hypersecretion of gastric acid, causing hypersecretion of pancreatic exocrine secretion, dilating blood vessels, increasing cardiac contractility, and protecting cardiomyocytes. A method or non-therapeutic method for any one of the group consisting of promoting growth hormone secretion, promoting fat accumulation (inhibiting weight loss), inhibiting insulin secretion, inhibiting the function of brown adipose tissue, promoting digestive tract motility, hypersecretion of gastric acid, hypersecretion of pancreatic exocrine secretion, dilating blood vessels, increasing cardiac contractility, and protecting cardiomyocytes, comprising the step of administering a composition containing bacteria belonging to Lactobacillus delbrueckii to a subject. A use or non-therapeutic use of a composition containing bacteria belonging to Lactobacillus delbrueckii, selected from the group consisting of promoting growth hormone secretion, promoting fat accumulation (inhibiting weight loss), inhibiting insulin secretion, inhibiting the function of brown adipose tissue, promoting digestive tract motility, hypersecretion of gastric acid, hypersecretion of pancreatic exocrine secretion, dilating blood vessels, increasing cardiac contractility, and protecting cardiomyocytes.

[0054]

[11] The composition, use in manufacturing, method or non-therapeutic method, or use or non-therapeutic use according to

[10] , wherein any one of the group consisting of promoting growth hormone secretion, promoting fat accumulation (inhibiting weight loss), inhibiting insulin secretion, inhibiting the function of brown adipose tissue, promoting gastrointestinal motility, increasing gastric acid secretion, increasing pancreatic exocrine secretion, dilating blood vessels, increasing cardiac contractility and protecting cardiomyocytes is based on promoting gastrointestinal secretion.

[0055]

[12] The composition, use in manufacture, method or non-therapeutic method, or use or non-therapeutic use according to any one of [9] to

[11] , wherein the bacterium belonging to Lactobacillus delbrueckii is a bacterium belonging to any one of Lactobacillus delbrueckii ssp. bulgaricus, Lactobacillus delbrueckii ssp. delbrueckii, Lactobacillus delbrueckii ssp. indicus, Lactobacillus delbrueckii ssp. lactis and Lactobacillus delbrueckii ssp. sunkii.

[0056]

[13] The composition, use in manufacture, method or non-therapeutic method, or use or non-therapeutic use according to any one of [9] to

[12] , wherein the bacterium belonging to Lactobacillus delbrueckii subsp. bulgaricus is selected from Lactobacillus delbrueckii subsp. bulgaricus 2038, Lactobacillus delbrueckii subsp. bulgaricus OLL1255 (Accession No.: NITE BP-76), Lactobacillus delbrueckii subsp. bulgaricus OLL1171 (Accession No.: NITE BP-01569), Lactobacillus delbrueckii subsp. bulgaricus OLL1247 (Accession No.: NITE BP-01814), Lactobacillus delbrueckii subsp. bulgaricus OLL205013 (Accession No.: NITE BP-02411), Lactobacillus delbrueckii OLL204989 (Accession No.: NITE BP-02874), Lactobacillus delbrueckii subsp. delbrueckii (JCM 1012T), Lactobacillus delbrueckii subsp. indica (JCM Any one of the group consisting of Lactobacillus delbrueckii subsp. lactis (JCM 1248T) and Lactobacillus delbrueckii subsp. mori (JCM17838T).

[0057]

[14] The composition, use in production, method or non-therapeutic method, or use or non-therapeutic use according to any one of [9] to

[13] , wherein the bacteria belonging to Lactobacillus delbrueckii are live bacteria.

[0058]

[15] The composition, use in production, method or non-therapeutic method, or use or non-therapeutic use according to any one of [9] to

[14] , wherein the composition is fermented milk that does not contain bacteria belonging to Lactobacillus paragaseri.

[0059]

[16] The composition, use in production, method or non-therapeutic method, or use or non-therapeutic use according to any one of [1] to [7], wherein the bacteria belonging to Lactobacillus delbrueckii does not have a DNA consisting of the sequence of SEQ ID NO: 6.

[0060] Effects of the Invention

[0061] According to the present invention, the secretion of ghrelin can be promoted by using a highly safe ingredient. BRIEF DESCRIPTION OF THE DRAWINGS

[0062] Figure 1 The results of measuring the mRNA expression of somatolin in MKN45 cells after incubation of each strain of Lactobacillus delbrueckii subsp. bulgaricus for 4 hours or 6 hours are shown. Mean ± SE, n = 3, * p<0.05, ** p<0.01vs. control (based on Dunnett's test)

[0063] Figure 2 The results of measuring the mRNA expression of ghrelin when using Lactobacillus delbrueckii OLL204989, Lactobacillus delbrueckii subsp. delbrueckii JCM 1012T, Lactobacillus delbrueckii subsp. indica JCM 15610T, Lactobacillus delbrueckii subsp. lactis JCM 1248T, or Lactobacillus delbrueckii subsp. mori JCM 17838T are shown. Mean ± SE, n = 3, * p<0.05, ** p<0.01vs. control (based on Dunnett's test) DETAILED DESCRIPTION

[0064] The present invention is described in detail below.

[0065] In one embodiment, the present invention relates to a composition containing specific bacteria. More specifically, it relates to a composition for promoting ghrelin secretion, etc., containing specific bacteria as an active ingredient. It should be noted that in the present invention, when the term "arbitrary" is used, it means that the number and type are arbitrary.

[0066] [Active ingredients]

[0067] "Bacteria belonging to the group of Lactobacillus delbrueckii"

[0068] The composition of the present invention contains bacteria belonging to Lactobacillus delbrueckii (hereinafter sometimes referred to as L. delbrueckii) as an active ingredient. In the composition, any bacteria belonging to L. delbrueckii can be used as an active ingredient regardless of specific subspecies.

[0069] The taxonomically equivalent strains to a certain strain (hereinafter referred to as strain S) are, for example, any of the following.

[0070] A strain that belongs to the same genus as strain S, preferably the same species as strain S, and whose entire sequence or a characteristic portion of its 16S rRNA gene (V1 region, V2 region, or all or a portion of V1 region and V2 region, or a portion including V1 region and V2 region, etc.) has a sequence identity of 90% or more, preferably 95% or more, more preferably 98% or more, further preferably 98.5% or more, further preferably more than 98.7%, further preferably 99% or more, further preferably 100% with the sequence of strain S;

[0071] · A strain with the same mycological properties as strain S.

[0072] It should be noted that regarding the criteria for determining the similarity and difference of species based on 16S rRNA gene sequences, those skilled in the art may refer to Stackebrandt E, Ebers J. Taxonomic parameters revisited: tarnished gold standards. Microbiol Today 2006; 33: 152-155.

[0073] (L.delbrueckii OLL204989)

[0074] As an example of strains belonging to L. delbrueckii, OLL204989 (deposit number: NITE BP-02874) and its taxonomically equivalent strains are used. L. delbrueckii OLL204989 was internationally deposited with the Patent Organism Depository of the Institute of Technology for Evaluation of Products (2-5-8, Kazusa Kamata, Kisarazu, Chiba Prefecture, Japan) under the deposit number NITE BP-02874 (depositor: Meiji Co., Ltd., deposit date of international deposit based on the Budapest Treaty: February 5, 2019).

[0075] <Bacteria belonging to Lactobacillus delbrueckii subspecies bulgaricus>

[0076] In a preferred embodiment, the bacteria belonging to L. delbrueckii in the composition of the present invention are bacteria belonging to Lactobacillus delbrueckii subspecies bulgaricus (Lactobacillus delbrueckii subspecies bulgaricus (hereinafter sometimes referred to as L. delbrueckii ssp. bulgaricus.)). L. delbrueckii ssp. bulgaricus is a lactic acid bacterium suitable for fermented milk, and the use of this bacterium facilitates the production of fermented milk. In the composition, as long as the bacteria belong to L. delbrueckii ssp. bulgaricus, it is not limited to a specific strain, and various bacteria can be used as active ingredients.

[0077] As examples of strains belonging to L. delbrueckii ssp. bulgaricus, 2038, OLL1255 (Accession No.: NITE BP-76), OLL1171 (Accession No.: NITE BP-01569), OLL1247 (Accession No.: NITE BP-01814), OLL205013 (Accession No.: NITE BP-02411), and taxonomically equivalent strains thereof are used.

[0078] (L.delbrueckii ssp.bulgaricus 2038)

[0079] L. delbrueckii ssp. bulgaricus 2038 can be isolated from Meiji Bulgarian Yogurt LB81 (Meiji Co., Ltd.) and can be obtained as a commercial product. In addition, it is also stored at Meiji Co., Ltd. (Meiji Technology Innovation Center, 1-29-1 Shichikoku, Hachioji City, Tokyo).

[0080] Alternatively, for example, 2038 strains can be isolated by applying a dilution of Meiji Bulgarian Yogurt LB81 (manufactured by Meiji Co., Ltd.) to an agar medium supplemented with BCP, culturing the mixture at 37°C for 48 hours, and then picking rough colonies.

[0081] The mycological properties of 2038 are shown below.

[0082] Morphology: Bacillus, does not produce gas from glucose, GC content 49.7%, Gram-positive, catalase-negative, lactic acid optical rotation: D type, does not grow at 15°C, can assimilate glucose, lactose, etc. to produce acid, does not form spores, and is non-motile.

[0083] The sequence of 2038 16S rRNA gene (corresponding to positions 27 to 520 of the full-length sequence of the rRNA gene) is shown in SEQ ID NO: 1 in the sequence listing.

[0084] 16S rRNA gene, Lactobacillus delbrueckii subsp. bulgaricus 2038 (SEQ ID NO. 1)

[0085] gctggcggcgtgcctaatacatgcaagtcgagcgagctgaattcaaagatyccttcgggrtgatttgttggacgctagcggcggatgggtgagtaacacgtgggcaatctgccctaaagactggg ataccacttggaaacaggtgctaataccggataacaacatgaatcgcatgattcaagtttgaaaggcggcgyaagctgtcactttaggatgagcccgcggcgcattagctagttggtggggtaaag gcctaccaaggcaatgatgcgtagccgagttgagagactgatcggccacattgggactgagacacggcccaaactcctacgggaggcagcagtagggaatcttccacaatggacgcaagtctgat ggagcaacgccgcgtgagtgaagaaggttttcggatcgtaaaagctctgttgttggtgaagaaggatagaggcagtaactggtctttatttgacggtaatcaaccagaaagtcacggctaactacgt

[0086] (L.delbrueckii ssp.bulgaricus OLL1255)

[0087] L.delbrueckii ssp.bulgaricus OLL1255 (accession number: NITE BP-76) was internationally deposited with the Patent Organism Depository of the Independent Administrative Institution for Product Evaluation and Technology (2-5-8, Kazusa Kamata, Kisarazu-shi, Chiba Prefecture, Japan) under the accession number NITEBP-76 (depositor: Meiji Co., Ltd., domestic deposit date: February 10, 2005, date of receipt of transfer request for international deposit based on the Budapest Treaty: April 1, 2009).

[0088] The mycological properties of OLL1255 are shown below.

[0089] Morphology: bacillus, non-gas-producing, Gram-positive, catalase-negative, facultative anaerobic, non-nitrate-reducing, non-indole-producing, non-hydrogen sulfide-producing, lactic acid. Optical rotation: D-type, no growth at 15°C, no spore formation, no motility.

[0090] Glycolytic properties: Arabinose-, Galactose-, Glucose+, Salicin-, Sucrose-, Cellobiose-, Starch-, Maltose-, Mannitol-, Mannitol+, Lactose+, Raffinose-, Ribose-.

[0091] The sequence of the 16S rRNA gene of OLL1255 (corresponding to positions 27 to 520 of the full-length sequence of the rRNA gene) is shown in SEQ ID NO: 2 in the sequence listing.

[0092] 16S rRNA gene, Lactobacillus delbrueckii subsp. bulgaricus OLL1255 (SEQ ID NO: 2)

[0093] gctggcggcgtgcctaatacatgcaagtcgagcgagctgaattcaaagatyccttcgggrtgatttgttggacgctagcggcggatgggtgagtaacacgtgggcaatctgccctaaagactggg ataccacttggaaacaggtgctaataccggataacaacatgaatcgcatgattcaagtttgaaaggcggcgyaagctgtcactttaggatgagcccgcggcgcattagctagttggtggggtaaag gcctaccaaggcaatgatgcgtagccgagttgagagactgatcggccacattgggactgagacacggcccaaactcctacgggaggcagcagtagggaatcttccacaatggacgcaagtctgat ggagcaacgccgcgtgagtgaagaaggttttcggatcgtaaaagctctgttgttggtgaagaaggatagaggcagtaactggtctttatttgacggtaatcaaccagaaagtcacggctaactacgt

[0094] (L.delbrueckii ssp.bulgaricus OLL1171)

[0095] L.delbrueckii ssp.bulgaricus OLL1171 (deposit number: NITE BP-01569) was internationally deposited with the Patent Organism Depository of the Independent Administrative Institution, National Institute of Technology for Product Evaluation (2-5-8, Kazusa Kamata, Kisarazu-shi, Chiba Prefecture, Japan) under the deposit number NITE BP-01569 (depositor: Meiji Co., Ltd., deposit date of international deposit based on the Budapest Treaty: March 13, 2013).

[0096] The sequence of the 16S rRNA gene of OLL1171 (corresponding to positions 27 to 520 of the full-length sequence of the rRNA gene) is shown in SEQ ID NO: 3 in the sequence listing.

[0097] 16S rRNA gene, Lactobacillus delbrueckii subsp. bulgaricus OLL1171 (SEQ ID NO: 3)

[0098] gctggcggcgtgcctaatacatgcaagtcgagcgagctgaattcaaagatyccttcgggrtgatttgttggacgctagcggcggatgggtgagtaacacgtgggcaatctgccctaaagactggg ataccacttggaaacaggtgctaataccggataacaacatgaatcgcatgattcaagtttgaaaggcggcgyaagctgtcactttaggatgagcccgcggcgcattagctagttggtggggtaaag gcctaccaaggcaatgatgcgtagccgagttgagagactgatcggccacattgggactgagacacggcccaaactcctacgggaggcagcagtagggaatcttccacaatggacgcaagtctgat ggagcaacgccgcgtgagtgaagaaggttttcggatcgtaaaagctctgttgttggtgaagaaggatagaggcagtaactggtctttatttgacggtaatcaaccagaaagtcacggctaactacgt

[0099] (L.delbrueckii ssp.bulgaricus OLL1247)

[0100] L.delbrueckii ssp.bulgaricus OLL1247 (deposit number: NITE BP-01814) was internationally deposited with the Patent Organism Depository of the Independent Administrative Institution, National Institute of Technology for Product Evaluation (2-5-8, Kazusa Kamata, Kisarazu-shi, Chiba Prefecture, Japan) under the deposit number NITE BP-01814 (depositor: Meiji Co., Ltd., deposit date of international deposit based on the Budapest Treaty: March 6, 2014).

[0101] The sequence of the 16S rRNA gene of OLL1247 (corresponding to positions 27 to 520 of the full-length sequence of the rRNA gene) is shown in SEQ ID NO: 4 in the sequence listing.

[0102] 16S rRNA gene, Lactobacillus delbrueckii subsp. bulgaricus OLL1247 (SEQ ID NO: 4)

[0103] gctggcggcgtgcctaatacatgcaagtcgagcgagctgaattcaaagatyccttcgggrtgatttgttggacgctagcggcggatgggtgagtaacacgtgggcaatctgccctaaagactggg ataccacttggaaacaggtgctaataccggataacaacatgaatcgcatgattcaagtttgaaaggcggcgyaagctgtcactttaggatgagcccgcggcgcattagctagttggtggggtaaag gcctaccaaggcaatgatgcgtagccgagttgagagactgatcggccacattgggactgagacacggcccaaactcctacgggaggcagcagtagggaatcttccacaatggacgcaagtctgat ggagcaacgccgcgtgagtgaagaaggttttcggatcgtaaaagctctgttgttggtgaagaaggatagaggcagtaactggtctttatttgacggtaatcaaccagaaagtcacggctaactacgt

[0104] (L.delbrueckii ssp.bulgaricus OLL205013)

[0105] L.delbrueckii ssp.bulgaricus OLL205013 (deposit number: NITE BP-02411) was internationally deposited with the Patent Organism Depository of the Independent Administrative Institution, National Institute of Technology for Product Evaluation (2-5-8, Kazusa Kamata, Kisarazu-shi, Chiba Prefecture, Japan) under the deposit number NITE BP-02411 (depositor: Meiji Co., Ltd., deposit date of international deposit based on the Budapest Treaty: February 13, 2017).

[0106] The sequence of the 16S rRNA gene of OLL205013 (corresponding to positions 27 to 520 in the full-length sequence of the rRNA gene) is shown in SEQ ID NO: 5 in the sequence listing.

[0107] 16S rRNA gene, Lactobacillus delbrueckii subsp. bulgaricus OLL205013 (SEQ ID NO: 5)

[0108] gctggcggcgtgcctaatacatgcaagtcgagcgagctgaattcaaagatyccttcgggrtgatttgttggacgctagcggcggatgggtgagtaacacgtgggcaatctgccctaaagactggg ataccacttggaaacaggtgctaataccggataacaacatgaatcgcatgattcaagtttgaaaggcggcgyaagctgtcactttaggatgagcccgcggcgcattagctagttggtggggtaaag gcctaccaaggcaatgatgcgtagccgagttgagagactgatcggccacattgggactgagacacggcccaaactcctacgggaggcagcagtagggaatcttccacaatggacgcaagtctgat ggagcaacgccgcgtgagtgaagaaggttttcggatcgtaaaagctctgttgttggtgaagaaggatagaggcagtaactggtctttatttgacggtaatcaaccagaaagtcacggctaactacgt

[0109] <Bacteria belonging to Lactobacillus delbrueckii subspecies delbrueckii>

[0110] In a preferred embodiment, the bacteria belonging to L. delbrueckii in the composition of the present invention are bacteria belonging to Lactobacillus delbrueckii ssp. delbrueckii (hereinafter sometimes referred to as L. delbrueckii ssp. delbrueckii). In the composition, the bacteria belonging to L. delbrueckii ssp. delbrueckii are not limited to specific strains, and various bacteria can be used as active ingredients.

[0111] (L.delbrueckii ssp.delbrueckii JCM 1012 T )

[0112] As an example of a strain belonging to L. delbrueckii ssp. delbrueckii, JCM 1012 can be used. T and its taxonomically equivalent strains. L.delbrueckii ssp.delbrueckii JCM 1012 T The PCR product can be obtained from the Japan Collection of Microorganisms, RIKEN BRC-JCM, Japan, under the JCM number JCM 1012. T And obtain.

[0113] <Bacteria belonging to Lactobacillus delbrueckii subspecies indica>

[0114] In a preferred embodiment, the bacteria belonging to L. delbrueckii in the composition of the present invention are bacteria belonging to Lactobacillus delbrueckii ssp. indicus (hereinafter sometimes referred to as L. delbrueckii ssp. indicus). In the composition, the bacteria belonging to L. delbrueckii ssp. indicus are not limited to specific strains, and various bacteria can be used as active ingredients.

[0115] (L. delbrueckii ssp. indicus JCM 15610 T )

[0116] As an example of a strain belonging to L. delbrueckii ssp. indicus, JCM 15610 was used. T and its taxonomically equivalent strains. L.delbrueckii ssp.indicus JCM 15610 T The PCR product can be obtained from the Japan Collection of Microorganisms, RIKEN BRC (RIKEN BRC-JCM, Japan) under the JCM number JCM 15610. T Obtain.

[0117] <Bacteria belonging to Lactobacillus delbrueckii subsp. lactis>

[0118] In a preferred embodiment, the bacteria belonging to L. delbrueckii in the composition of the present invention are bacteria belonging to Lactobacillus delbrueckii ssp. lactis (hereinafter sometimes referred to as L. delbrueckii ssp. lactis). In the composition, the bacteria belonging to L. delbrueckii ssp. lactis are not limited to specific strains, and various bacteria can be used as active ingredients.

[0119] (L.delbrueckii ssp.lactis JCM 1248 T )

[0120] As an example of a strain belonging to L. delbrueckii ssp. lactis, JCM 1248 was used. T and its taxonomically equivalent strains. L.delbrueckii ssp.lactis JCM 1248 T The PCR product can be obtained from the Japan Collection of Microorganisms, RIKEN BRC (RIKEN BRC-JCM, Japan) under the JCM number JCM 1248. T Obtain.

[0121] <Bacteria belonging to Lactobacillus delbrueckii subspecies Morus albus>

[0122] In a preferred embodiment, the bacteria belonging to L. delbrueckii in the composition of the present invention are bacteria belonging to Lactobacillus delbrueckii ssp. sunkii (hereinafter sometimes referred to as L. delbrueckii ssp. sunkii). In the composition, the bacteria belonging to L. delbrueckii ssp. sunkii are not limited to specific strains, and various bacteria can be used as active ingredients.

[0123] (L. delbrueckii ssp. sunkii JCM 17838 T )

[0124] As an example of a strain belonging to L. delbrueckii ssp. sunkii, JCM 17838 was used. T and its taxonomically equivalent strains. L.delbrueckii ssp.sunkii JCM 17838 TThe PCR product can be obtained from the Japan Collection of Microorganisms, RIKEN BRC (RIKEN BRC-JCM, Japan) under the JCM number JCM 17838. T Obtain.

[0125] As described above, in a preferred embodiment, the bacteria belonging to L.delbrueckii in the composition of the present invention are bacteria belonging to any one of L.delbrueckii ssp., L.delbrueckii ssp.bulgaricus, L.delbrueckiissp.delbrueckii, L.delbrueckii ssp.indicus, L.delbrueckii ssp.lactis and L.delbrueckii ssp.sunkii. In another embodiment, the bacteria belonging to L.delbrueckii do not contain bacteria belonging to Lactobacillus delbrueckii subspecies bulgaricus. Furthermore, in another embodiment, the bacteria belonging to L.delbrueckii in the composition of the present invention are bacteria belonging to any one of L.delbrueckii ssp.delbrueckii, L.delbrueckiissp.indicus, L.delbrueckii ssp.lactis and L.delbrueckii ssp.sunkii.

[0126] In addition, the bacteria belonging to L. delbrueckii in the composition of the present invention may be selected from L. delbrueckii ssp. bulgaricus 2038, L. delbrueckii ssp. bulgaricus OLL1255 (Accession No.: NITE BP-76), L. delbrueckii ssp. bulgaricus OLL1171 (Accession No.: NITE BP-01569), L. delbrueckii ssp. bulgaricus OLL1247 (Accession No.: NITE BP-01814), L. delbrueckii ssp. bulgaricus OLL205013 (Accession No.: NITE BP-02411), L. delbrueckii OLL204989 (Accession No.: NITE BP-02874), L. delbrueckii ssp. delbrueckii (JCM 1012T), L. delbrueckii ssp.indicus (JCM15610T), L.delbrueckii ssp.lactis (JCM 1248T) and L.delbrueckii ssp.sunkii (JCM17838T) The group consisting of any one or more. In another embodiment, the bacteria belonging to L.delbrueckii in the composition of the present invention does not contain L.delbrueckii LAB4. Furthermore, in another embodiment, the bacteria belonging to L.delbrueckii in the composition of the present invention are selected from any one or more of the group consisting of L.delbrueckii ssp.delbrueckii (JCM 1012T), L.delbrueckii ssp.indicus (JCM 15610T), L.delbrueckii ssp.lactis (JCM 1248T) and L.delbrueckii ssp.sunkii (JCM 17838T).

[0127] In another embodiment, the bacteria belonging to L. delbrueckii in the composition of the present invention have a 16S rRNA gene that is 90% or more, preferably 95% or more, more preferably 98% or more, further preferably 98.5% or more, further preferably 98.7% or more, particularly preferably 99% or more, and preferably 99.5% or more identical to the base sequence represented by any one of SEQ ID NOs: 1 to 5.

[0128] A composition comprising any of the above-mentioned strains as an active ingredient is particularly excellent in the target effect and is preferred.

[0129] <DNA encoding hypothetical protein gene>

[0130] The present inventors have found that one of the common characteristics of the strains that have been confirmed to have a ghrelin secretion-promoting effect under the conditions of this embodiment is that they do not have the DNA (DNA encoding a hypothetical protein gene) with the sequence recorded below as sequence number 6. The amino acid sequence of the protein (hypothetical protein) corresponding to the DNA is recorded in sequence number 7 of the sequence table. Therefore, in a preferred embodiment, the composition may contain any bacteria belonging to Lactobacillus delbrueckii and not having the following DNA (i) as an active ingredient. In other words, in this embodiment, the bacteria belonging to Lactobacillus delbrueckii contained in the composition do not have the DNA of (i) or do not contain strains with the DNA of (i). In another embodiment, the composition may contain any bacteria belonging to Lactobacillus delbrueckii and not having the following DNA (ii) as an active ingredient. Furthermore, in another embodiment, the composition may contain any bacteria belonging to Lactobacillus delbrueckii and not having the following DNA (iii) as an active ingredient.

[0131] (i) DNA consisting of the sequence of SEQ ID NO: 6

[0132] (ii) a DNA comprising a sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to SEQ ID NO: 6

[0133] (iii) a DNA comprising a sequence in which 1, 1 to 2, 1 to 3, 1 to 4, 1 to 5, 1 to 6, 1 to 7, 1 to 8, 1 to 9, 1 to 10, 1 to 11, 1 to 12, 1 to 13, 1 to 14, 1 to 15, 1 to 16, 1 to 17, 1 to 18, 1 to 19, 1 to 20 or 1 to 21 bases are substituted, deleted or added in SEQ ID NO: 6

[0134] In addition, in another preferred embodiment, the composition may include any bacteria belonging to Lactobacillus delbrueckii and not having the following protein (iv) as an active ingredient. In other words, in this embodiment, the bacteria belonging to Lactobacillus delbrueckii contained in the composition do not have the protein (iv) or do not contain a strain having the protein (iv). In another embodiment, the composition may include any bacteria belonging to Lactobacillus delbrueckii and not having the following protein (v) as an active ingredient. Furthermore, in another embodiment, the composition may include any bacteria belonging to Lactobacillus delbrueckii and not having the following protein (vi) as an active ingredient.

[0135] (iv) Protein consisting of the sequence of SEQ ID NO: 7

[0136] (v) a protein encoded by the DNA of (ii) above

[0137] (vi) a protein encoded by the DNA of (iii) above

[0138] Furthermore, in another embodiment, the composition may contain any bacteria that does not have a DNA encoding a protein consisting of the sequence of SEQ ID NO: 7 (may contain a DNA consisting of the sequence of SEQ ID NO: 6). In other words, in this embodiment, the bacteria belonging to Lactobacillus delbrueckii contained in the composition do not have a DNA encoding a protein consisting of the sequence of SEQ ID NO: 7 or do not contain a strain having a DNA encoding a protein consisting of the sequence of SEQ ID NO: 7. In another preferred embodiment, the composition may contain any bacteria belonging to Lactobacillus delbrueckii subspecies bulgaricus and not having a DNA consisting of the sequence of SEQ ID NO: 6 as an active ingredient. In another preferred embodiment, the composition may contain any bacteria belonging to Lactobacillus delbrueckii subspecies bulgaricus and not having a protein consisting of the sequence of SEQ ID NO: 7 as an active ingredient.

[0139] DNA encoding hypothetical protein gene (SEQ ID NO: 6)

[0140] atggttgatgagatgtaccctgaagccgagaagatcattcttgtcatagacaatcttaatacgcacaagccatcctcactatataaggctttcaaagcacgtcgaatc atcaagaagcttgagattcactatactcccaagcacggcagctggttagatatcgctgaaattgatgatgaccagacagtgtctttctcgaagaatcccggatattga

[0141] hypothetical protein (SEQ ID NO: 7)

[0142] MVDEMYPEAEKIILVIDNLNTHKPSSLYKAFKARRIIKKLEIHYTPKHGSWLDIAEIDDDQTVSFSKNPGY

[0143] It should be noted that, in the composition, lactic acid bacteria other than bacteria belonging to L.delbrueckii may be included as long as the target function is not impaired. In addition, the composition may include microorganisms such as yeast other than the target lactic acid bacteria. In a preferred embodiment, the composition includes one or more lactic acid bacteria, and does not contain microorganisms other than them, such as yeast. In addition, in another preferred embodiment, the composition does not contain any one or both of Lactobacillus gasseri and Lactobacillus paragasseri. In addition, in another preferred embodiment, the composition does not contain Streptococcus thermophilus. In addition, in another preferred embodiment, the composition does not contain L.delbrueckiissp.bulgaricus OLL1073R-1. In addition, in another preferred embodiment, the composition does not contain lactic acid bacteria other than bacteria belonging to L.delbrueckii. In addition, in another preferred embodiment, the composition does not contain lactic acid bacteria other than bacteria belonging to L.delbrueckii. In another preferred embodiment, the composition does not contain any component that promotes the secretion of ghrelin other than bacteria belonging to L. delbrueckii. In another preferred embodiment, the composition does not contain any component that promotes the secretion of ghrelin other than bacteria belonging to L. delbrueckii ssp. bulgaricus.

[0144] The lactic acid bacteria (effective bacteria) as the effective ingredient contained in the composition can be produced by culturing. There are no particular restrictions on the culturing conditions as long as the target effect can be exerted.

[0145] In the composition of the present invention, the effective bacteria can be contained in any state as long as they can exert the target effect. For example, in addition to the effective bacteria itself, the effective bacteria may also be a culture of the effective bacteria (including the effective bacteria and the culture supernatant). The bacteria can be in a living state (live bacteria) or in a dead state (dead bacteria) as long as they can exert the target effect.

[0146] The effective bacteria may be contained in the composition in the form of dead bacteria. The dead bacteria may be obtained by sterilizing the effective bacteria. The sterilization treatment is not particularly limited as long as it can exert the target effect, and heating, sterilization lamp (UV), ozone, chemicals, high osmotic pressure, etc. may be used.

[0147] The dead bacteria are preferably heat-treated dead bacteria obtained by heat-treating live bacteria. The heat treatment used to obtain the heat-treated dead bacteria is not particularly limited as long as the target effect can be exerted, and can be implemented at a temperature and time sufficient to kill the effective bacteria used. Such conditions also depend on the effective bacteria used, but the heat treatment temperature is, for example, above 55°C, preferably above 60°C, more preferably above 65°C, further preferably above 70°C, can be set to above 80°C, and can be set to above 90°C. The upper limit of the heat treatment temperature can be appropriately set, for example, below 121°C, below 100°C, below 90°C, and below 80°C. In addition, according to the heat treatment temperature, the heat treatment time can be set to more than 1 minute, can be set to more than 3 minutes, can be set to more than 10 minutes, more than 15 minutes, more than 30 minutes, and more than 45 minutes. The upper limit of the heat treatment time can be set, for example, below 120 minutes, can be below 100 minutes, can be below 90 minutes, and can be below 80 minutes.

[0148] The live cells can be obtained by culturing the live cells by a known method, and can be in the form of a culture solution, or in the form of a concentrate, frozen product, or dried product of the culture solution. Alternatively, a substance obtained by subjecting the culture solution to centrifugal separation or the like and recovering only the cells, or a substance obtained by suspending the recovered cells in a buffer solution can also be used. Alternatively, live cells are added (inoculated) to food materials that are raw materials for fermented foods, such as milk (animal milk), to increase the amount of live cells in the composition endogenously.

[0149] The effective bacteria to be included in the composition, whether living bacteria or dead bacteria, can be prepared in the form of dry matter, suspension, paste, gel, etc.

[0150] [use]

[0151] (Function)

[0152] In one embodiment, the composition can be used to promote ghrelin secretion. Gastrointestinal hormone is a peptide hormone that is mainly secreted in the stomach. In addition, the composition can also be used to help, assist, improve or maintain ghrelin secretion. In addition, the composition can also be used to alleviate the decrease in ghrelin secretion, reduce the risk of decreased ghrelin secretion, or help improve ghrelin secretion.

[0153] Gastrointestinal growth hormone is known to promote growth hormone secretion (Patent Documents 1, 2), inhibit the function of brown adipose tissue, promote digestive tract motility, hypersecretion of gastric acid, hypersecretion of pancreatic exocrine, dilate blood vessels, increase cardiac contractility and protect myocardial cells (Patent Document 3) and other effects. Therefore, in one embodiment, the composition can be used to select any one or more of the group consisting of promoting growth hormone secretion, inhibiting the function of brown adipose tissue, promoting digestive tract motility (or helping digestion), hypersecretion of gastric acid, hypersecretion of pancreatic exocrine, dilate blood vessels, increase cardiac contractility and protect myocardial cells. Alternatively, the composition can be used to regulate or help regulate any one or more of the group consisting of promoting growth hormone secretion, inhibiting the function of brown adipose tissue, promoting digestive tract motility, hypersecretion of gastric acid, hypersecretion of pancreatic exocrine, dilate blood vessels, increase cardiac contractility and protect myocardial cells.

[0154] In addition, it is known that ghrelin has the effects of promoting fat accumulation (inhibiting weight loss) and inhibiting insulin secretion in addition to the above-mentioned effects (patent document 3). Therefore, in one embodiment, the composition can be used to select any one or more of the group consisting of promoting growth hormone secretion, promoting fat accumulation (inhibiting weight loss), inhibiting insulin secretion, inhibiting the function of brown adipose tissue, promoting digestive tract motility (or helping digestion), hypersecretion of gastric acid, hypersecretion of pancreatic exocrine, dilating blood vessels, increasing cardiac contractility and protecting myocardial cells. Alternatively, the composition can be used to regulate or help regulate any one or more of the group consisting of promoting growth hormone secretion, promoting fat accumulation (inhibiting weight loss), inhibiting insulin secretion, inhibiting the function of brown adipose tissue, promoting digestive tract motility, hypersecretion of gastric acid, hypersecretion of pancreatic exocrine, dilating blood vessels, increasing cardiac contractility and protecting myocardial cells.

[0155] In addition, the following uses of the composition of the present invention may be excluded: regulating the autonomic nervous system, inhibiting the sympathetic nervous system, improving cold symptoms, improving stiffness, improving gastrointestinal discomfort, improving worsening depression, improving worsening dizziness, improving increased heart rate, improving increased blood pressure, improving worsening chronic pain, improving fatigue, improving general fatigue, improving general lethargy, improving conscious stress, improving daytime sleepiness, improving anxiety, improving loss of appetite, improving decreased thinking ability, improving dry mouth, improving loss of refreshment, promoting fat accumulation (inhibiting weight loss), inhibiting weight loss associated with ulcerative colitis, inhibiting insulin secretion, and inhibiting excessive insulin secretion after sugar intake.

[0156] Whether a certain component can promote the secretion of ghrelin can be confirmed by various methods by those skilled in the art. For example, the component can be added to human gastric cells such as MKN45 cells (human gastric cancer cell line), and the expression level of the ghrelin gene in the cells can be measured after an appropriate period of time for evaluation. The expression level of the ghrelin gene can be specifically measured in the form of mRNA expression level. It is preferred to use the expression level of a constitutively expressed gene, such as β-actin, for correction.

[0157] (Target)

[0158] In one embodiment, the composition is suitable for ingestion by a subject who needs or wishes to promote the secretion of ghrelin. In one embodiment, the composition is suitable for ingestion by a subject whose secretion of ghrelin has decreased. In addition, in one embodiment, the composition is suitable for ingestion by a subject who needs or wishes to promote the secretion of growth hormone, promote fat accumulation (inhibit weight loss), inhibit insulin secretion, inhibit the function of brown adipose tissue, promote gastrointestinal motility, hypersecretion of gastric acid, hypersecretion of pancreatic exocrine secretion, dilate blood vessels, increase cardiac contractility and protect myocardial cells. The judgment of the above-mentioned disease or state of the subject also includes: the judgment of a physician, nurse, pharmacist, etc.; living habits and eating habits, output results of subjective symptom questionnaires, self-judgment based on subjective symptoms, etc. The subject can be healthy (for example, a healthy stomach, no disease to be treated, and no recipient of therapeutic drugs, such as chemotherapy agents). In addition, the following can be excluded from the subjects who take the composition of the present invention: patients with autonomic nervous system disorders, patients with cold symptoms, patients with stiffness, patients with depression, patients with dizziness, patients with summer heat symptoms, and patients with ulcerative colitis.

[0159] The above-mentioned subjects include adults (over 15 years old), middle-aged and elderly people, elderly people (over 65 years old (defined by the World Health Organization (WHO))), patients in the late stage of illness, pregnant women, parturients, infants and children. In addition, the active ingredient of the composition of the present invention is a bacterium with rich experience of use, so it is suitable for long-term intake, and is particularly suitable for subjects who wish to promote the secretion of ghrelin in daily life.

[0160] [Composition]

[0161] (Food composition, etc.)

[0162] The composition of the present invention can be made into a food composition or a pharmaceutical composition. Food and medicine include not only those for humans, but also those for animals other than humans, unless otherwise specified. Food includes conventional food, functional food, nutritional composition, and therapeutic food (food for therapeutic purposes. Food cooked based on a menu prepared by a dietitian, etc., based on a dietary prescription prescribed by a doctor), diet therapy food, formula food, nursing food, and therapeutic auxiliary food. Unless otherwise specified, food includes not only solids, but also liquid foods, such as beverages, drinking agents, liquid foods, and soups. Functional food refers to food that can impart a specified functionality to an organism, including, for example: specific health foods (including special protection [specific health foods] with additional conditions), functional labeled foods, health functional foods including nutritional functional foods, special purpose foods, nutritional supplements, health supplements, supplements (for example, tablets, coated tablets, sugar-coated tablets, capsules, liquids, etc.), beauty foods (for example, weight loss foods), and all other health foods. In addition, the "functional food" in the present invention includes health foods with health claims to which the food standards of the Codex Alimentarius Commission (FAO / WHO Joint Food Standards Commission) are applied.

[0163] The following substances are excluded from the compositions of the present invention:

[0164] A preparation containing protein hydrolysate and fermented milk protein as proteins, medium-chain fatty acids as lipids, and carbohydrates, wherein the fermented milk protein is derived from fermented milk obtained by fermenting skim milk using Lactobacillus Bulgaricus, Streptococcus thermophilus, or a combination thereof;

[0165] A composition comprising Lactobacillus casei, Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus delbrueckii subsp. bulgaricus, Streptococcus thermophilus, Bifidobacterium longum, Bifidobacterium breve, and Bifidobacterium infantis.

[0166] (Giving method)

[0167] The composition of the present invention may be administered parenterally, for example, via a tube (gastrostomy fistula, enteral fistula), nasally, or orally, and is preferably administered orally.

[0168] (Content and dosage of active ingredients)

[0169] The content of effective bacteria in the composition of the present invention can be any amount that can exert the target effect. The content of effective bacteria in the composition can be appropriately set in consideration of various factors such as the age, weight, and symptoms of the subject to be ingested or administered. One unit of the composition can be an amount that can be ingested or administered for one day or once (it can be administered or ingested once a day, or it can be administered multiple times a day, for example, three times).

[0170] In one embodiment, the content of effective bacteria per unit of the composition can be set to, for example, 1×10 5 More than 1×10 6 More than 1×10 7 More than 1×10 8 More than 5×10 8 More than 5.7×10 8 More than 6×10 8 More than 7×10 8 More than 8×10 8 More than 9×10 8 More than 1×10 9 More than 5×10 9 More than 1×10 10 More than 1×10 11 More than 2×10 11 The upper limit can be set appropriately, and the lower limit can be set to, for example, 1×10 14 Less than 1×10 13 Less than 1×10 12 Less than .

[0171] In one embodiment, the effective bacteria intake can be set to 1×10 5 For more than 10 6 For more than 10 7 For more than 10 8 More than 5×10 8 More than 5.7 × 10 8 More than 6×10 8 More than 7×10 8 More than 8×10 8 More than 9×10 8 More than 1×10 9 More than 5×10 9 More than 1×10 10 More than 1.3×1010 More than 2×10 10 More than 1×10 11 More than 2×10 11 The upper limit can be set appropriately, and the lower limit can be set to, for example, 1×10 14 If the number is less than 10 / day, it can be set to 1×10 13 If the number is less than 1 / day, it can be set to 1×10 12 Less than pcs / day.

[0172] It should be noted that commercially available yogurt sometimes contains 1×10 7 / g or more of L. delbrueckiissp.bulgaricus, the calculation result is that each product contains 1×10 9 More than one L.delbrueckiissp.bulgaricus.

[0173] In addition, when the content of effective bacteria is set as above, in any case, 1 unit can be set to, for example, 0.5g or more, 1g or more, 5g or more, 10g or more, 20g or more, 30g or more. When the composition is set in a food form such as fermented milk, as a food, 1 unit can be set to an amount that is easy to take in once, for example, 50g or more, 60g or more, 70g or more, 80g or more, 90g or more, 100g or more. The upper limit can be set appropriately, and the lower limit can be set to 500g or less in any case, and can be set to 400g or less, 300g or less, 200g or less, 150g or less, 125g or less.

[0174] The composition has effective bacteria that are well-known for consumption as an effective ingredient. Therefore, the composition of the present invention can be taken repeatedly or for a long time, for example, it can be continuously administered / taken for more than 3 days, preferably more than 1 week, more preferably more than 15 days, further preferably more than 3 weeks, more than 4 weeks, or more than 1 month.

[0175] (Other ingredients, additives)

[0176] The composition of the present invention may contain other effective ingredients and nutrients that are allowed as food or medicine. Examples of such ingredients are amino acids (e.g., lysine, arginine, glycine, alanine, glutamic acid, leucine, isoleucine, valine), sugars (glucose, sucrose, fructose, maltose, trehalose, erythritol, maltitol, palatinose, xylitol, dextrin), electrolytes (e.g., sodium, potassium, calcium, magnesium), vitamins (e.g., vitamin A, vitamin B1, vitamin B2, vitamin B6, vitamin B12, vitamin C, vitamin D, vitamin E, vitamin K, biotin, folic acid, pantothenic acid and nicotinic acid), minerals (e.g., copper, zinc, iron, cobalt, manganese), antibiotics, dietary fiber, protein, lipids, etc.

[0177] In addition, the composition may further comprise additives that are permitted as food or medicine. Examples of such additives are inactive carriers (solid carriers, liquid carriers), excipients, surfactants, binders, disintegrants, lubricants, solubilizers, suspending agents, coating agents, colorants, preservatives, buffers, pH regulators, emulsifiers, stabilizers, sweeteners, antioxidants, spices, acidulants, and natural substances. More specifically, water, other aqueous solvents, pharmaceutically acceptable organic solvents, collagen, polyvinyl alcohol, polyvinyl pyrrolidone, carboxyvinyl polymer, sodium alginate, water-soluble dextran, water-soluble dextrin, sodium carboxymethyl starch, pectin, xanthan gum, gum arabic, casein, gelatin, agar, glycerin, propylene glycol, polyethylene glycol, vaseline, paraffin, stearyl alcohol, stearic acid, human serum albumin, mannitol, sorbitol, lactose, sucralose, stevioside, aspartame, acesulfame potassium, citric acid, lactic acid, malic acid, tartaric acid, phosphoric acid, acetic acid, fruit juice, vegetable juice, etc.

[0178] (Dosage form and form)

[0179] The food composition of the present invention can be made into any form such as solid, liquid, mixture, suspension, powder, granule, paste, jelly, gel, capsule. In addition, the food composition of the present invention can be made into any form such as dairy products, supplements, desserts, beverages, supplements, seasonings, processed foods, side dishes, soups. More specifically, the composition of the present invention can be made into fermented milk, lactic acid bacteria beverage, milk beverage, milk beverage, refreshing beverage, ice cream, tablet, cheese, bread, biscuits (biscuits), crackers (cracker), pizza, formula milk powder, liquid food, patient food, nutritious food, frozen food, processed food and other forms, and can be made into forms such as granules, powders, pastes, thick liquids for mixing with beverages and food to take in addition. Fermented milk refers to the fermented milk and lactic acid bacteria beverage defined in "provincial decree on the composition standards of milk and dairy products (hereinafter referred to as "provincial decree such as milk".)". Fermented milk in the Ministry of Milk, etc. Ordinance refers to milk or milk containing equal to or more fat-free milk solids fermented with lactic acid bacteria or yeast and made into a paste or liquid, or frozen.

[0180] The pharmaceutical composition of the present invention can be prepared into any dosage form suitable for oral administration, such as solid preparations such as tablets, granules, powders, pills, capsules, liquid preparations such as liquids, suspensions, syrups, gels, aerosols, etc.

[0181] (other)

[0182] In the manufacture of the composition of the present invention, the stage of combining effective bacteria can be appropriately selected. As long as the characteristics of the effective bacteria are not significantly damaged, the stage of combination is not particularly limited. For example, the composition can be manufactured as follows: milk, dairy products and water are mixed, homogenized as needed, sterilized, inoculated with effective bacteria, and fermented, thereby manufacturing the composition in the form of fermented milk containing a sufficient amount of effective bacteria. The milk and dairy products as raw materials can be appropriately selected according to the effective bacteria used in the fermentation. In a preferred embodiment, the milk and dairy products as raw materials include milk and dairy products derived from animals. In addition, the milk and dairy products as raw materials may not contain vegetable milk, such as soy milk, almond milk, oat milk, coconut milk, rice milk, and hemp milk.

[0183] The composition can indicate the purpose of use (application), and can also indicate the content recommended for ingestion by a specific object. The specific object can be, for example, the object recorded in the above (object). The marking can be done directly or indirectly. Examples of direct marking include records on tangible objects such as the product itself, packaging, containers, labels, and tags. Examples of indirect marking include notifications through applications in AI diagnosis, web pages, stores, brochures, exhibitions, media seminars and other seminars, books, newspapers, magazines, television, radio, mail, email, audio, and other advertising and promotional activities based on places or means.

[0184] Examples of uses and functions indicated on the composition as a food include: promotion of ghrelin secretion, treatment of diseases or conditions that can be improved by promoting ghrelin secretion, such as promotion of growth hormone secretion, promotion of fat accumulation (inhibition of weight loss), inhibition of insulin secretion, inhibition of the function of brown adipose tissue, promotion of gastrointestinal motility, hypersecretion of gastric acid, hypersecretion of pancreatic exocrine secretion, dilation of blood vessels, increase of cardiac contractility and protection of cardiomyocytes, as well as the regulation, assistance and maintenance of these, use as probiotics, etc.

[0185] In one embodiment, the labeling of the recommended composition for ingestion is performed on an individual. Such labeling can be performed using a document (written or electronic) issued to the subject, a tablet terminal, a smartphone, a personal computer, or an SNS of the subject. In addition, such labeling can be performed together with the display of the results of any test or analysis such as a blood test or a blood metabolome test of the subject.

[0186] In one embodiment, a method for promoting ghrelin secretion in a subject is provided, comprising the following step: causing the subject to ingest a composition containing bacteria belonging to Lactobacillus delbrueckii based on the amount of ghrelin in the subject's blood (the blood concentration of ghrelin). In another embodiment, a method for promoting fat accumulation (suppressing weight loss) in a subject, a method for suppressing insulin secretion, a method for suppressing the function of brown adipose tissue, a method for promoting gastrointestinal motility, a method for hypersecretion of gastric acid, a method for hypersecretion of pancreatic exocrine, a method for dilating blood vessels, a method for increasing cardiac contractility, or a method for protecting cardiomyocytes is provided, comprising the following step: causing the subject to ingest a composition containing bacteria belonging to Lactobacillus delbrueckii based on the amount of ghrelin in the subject's blood (the blood concentration of ghrelin). Furthermore, in another embodiment, a method for providing a subject with a recommendation for ingestion of a composition, a method for supporting a subject's diet, or a method for supporting a subject's health is provided, comprising the following step: providing a recommendation for ingestion of a composition containing bacteria belonging to Lactobacillus delbrueckii based on the amount of ghrelin in the subject's blood (the blood concentration of ghrelin).

[0187] In these methods, the blood of the subject is preferably collected when fasting. The blood may be separated plasma. In addition, in these methods, the amount of ghrelin may be the amount of either or both of sialylated ghrelin (active form) and non-sialylated ghrelin. The amount of ghrelin can be measured by quantifying ghrelin in the blood, performing blood metabolome analysis, etc. The quantification of ghrelin can be performed using any method known to those skilled in the art, such as ELISA and HPLC. In addition, the subject in these methods may be, for example, a subject whose blood concentration of ghrelin when fasting is less than 5 pg / mL.

[0188] These methods may also include the following steps:

[0189] A step of collecting blood from a subject; a step of measuring the amount of ghrelin in the blood; a step of displaying the measurement result; a step of determining whether the subject desires or needs to promote the secretion of ghrelin based on the measurement result; a step of displaying the determination result; and a step of labeling a product as a composition containing bacteria belonging to Lactobacillus delbrueckii.

[0190] Example

[0191] [Example 1. Study on the effect of promoting ghrelin secretion by Lactobacillus delbrueckii subspecies bulgaricus]

[0192] MKN45 cells were cultured in RPMI-1640 medium without antibiotics on 24-well plates for 24 h.

[0193] The strains of Lactobacillus delbrueckii subsp. bulgaricus shown in Table 1 below were subjected to activation culture and main culture using MRS medium for 20 hours.

[0194] The cultured cells were washed with PBS and incubated with medium at OD 660 =0.5, and add the live bacteria to the plate. After 4 hours and 6 hours of incubation, the cells were washed 3 times with PBS, and 200μL of T / E solution and 200μL of Lysis Buffer of Maxwell RSC simplyRNA CellsKit (Promega) were added to recover the cells. Then, RNA was extracted and cDNA was converted, and the expression of the gastrin gene was measured by qPCR (SYBR Green-based embedding agent method). β-actin was used for standardization, and the analysis was performed by the ΔΔCt method as a comparative quantitative method.

[0195]

Table 1

[0196] Strains used and culture conditions

[0197]

[0198] ※It can be separated from Meiji Bulgarian Yogurt LB81 (Meiji Co., Ltd.).

[0199] The results are shown in Figure 1 Compared with the control group without live bacteria, the expression of ghrelin mRNA in the 2038 and OLL205013 groups increased significantly 4 hours after addition, and significantly increased in the P230250, OLL1255, OLL1171, and OLL1247 groups 6 hours later.

[0200] [Example 2. Study on the effect of promoting ghrelin secretion by various bacteria belonging to Lactobacillus delbrueckii]

[0201] The ghrelin secretion ability was studied by the same method as in Example 1 using the strains of Lactobacillus delbrueckii, Lactobacillus delbrueckii subsp. delbrueckii, Lactobacillus delbrueckii subsp. indica, Lactobacillus delbrueckii subsp. lactis, or Lactobacillus delbrueckii subsp. mori shown in Table 2 below.

[0202]

Table 2

[0203]

[0204] ※OLL204989 is not limited to subspecies.

[0205] The results are shown in Figure 2 Compared with the control group without live bacteria, ghrelin mRNA expression levels in all groups increased significantly 4 hours after addition, and significantly increased in all groups except the OLL204989 group 6 hours later.

[0206] As mentioned above, the strain belonging to L.delbrueckii promotes the expression of the gastrin gene, so it can be said that the bacteria belonging to L.delbrueckii can be used to promote the secretion of gastrin. In addition, the effects of gastrin such as promoting growth hormone secretion (patent documents 1, 2), promoting fat accumulation (inhibiting weight loss), inhibiting insulin secretion, inhibiting the function of brown adipose tissue, promoting digestive tract motility, causing hypersecretion of gastric acid, causing hypersecretion of pancreatic exocrine, dilating blood vessels, increasing cardiac contractility and protecting myocardial cells (patent document 3) are known. Therefore, it can be said that the bacteria belonging to L.delbrueckii can be used to select any one or more of the group consisting of promoting growth hormone secretion, promoting fat accumulation (inhibiting weight loss), inhibiting insulin secretion, inhibiting the function of brown adipose tissue, promoting digestive tract motility, causing hypersecretion of gastric acid, causing hypersecretion of pancreatic exocrine, dilating blood vessels, increasing cardiac contractility and protecting myocardial cells.

[0207] Industrial Applicability

[0208] The present invention can provide a food ingredient source with high safety or a material with confirmed safety as a ghrelin secretion promoter, thereby helping healthy people maintain and improve their health and improve their work efficiency. In addition, according to the present invention, a food composition and a method for producing a food useful for promoting ghrelin secretion or for maintaining and improving the health and improving the work efficiency of healthy people can be provided.

Claims

1. Use of a bacterium belonging to Lactobacillus delbrueckii for producing a composition for promoting ghrelin secretion.

2. Use of a bacterium belonging to Lactobacillus delbrueckii in the manufacture of a composition for promoting growth hormone secretion, inhibiting the function of brown adipose tissue, promoting digestive tract motility, increasing gastric acid secretion, increasing pancreatic exocrine secretion, dilating blood vessels, increasing cardiac contractility and protecting myocardial cells.

3. The use according to claim 2, wherein Any one or more selected from the group consisting of promoting growth hormone secretion, inhibiting the function of brown adipose tissue, promoting digestive tract motility, hypersecretion of gastric acid, hypersecretion of pancreatic exocrine, dilating blood vessels, increasing cardiac contractility and protecting myocardial cells is based on promoting the secretion of gastrin.

4. The use according to any one of claims 1 to 3, wherein The bacteria belonging to Lactobacillus delbrueckii are bacteria belonging to any one of Lactobacillus delbrueckii ssp. bulgaricus, Lactobacillus delbrueckii ssp. delbrueckii, Lactobacillus delbrueckii ssp. indicus, Lactobacillus delbrueckii ssp. lactis, and Lactobacillus delbrueckii ssp. sunkii.

5. The use according to any one of claims 1 to 3, wherein The bacteria belonging to Lactobacillus delbrueckii have a 16S rRNA gene having 90% or more identity with the base sequence represented by any one of SEQ ID NOs: 1 to 5.

6. The use according to any one of claims 1 to 3, wherein The bacteria belonging to Lactobacillus delbrueckii subsp. bulgaricus are selected from Lactobacillus delbrueckii subsp. bulgaricus 2038, Lactobacillus delbrueckii subsp. bulgaricus OLL1255 (Accession No.: NITE BP-76), Lactobacillus delbrueckii subsp. bulgaricus OLL1171 (Accession No.: NITE BP-01569), Lactobacillus delbrueckii subsp. bulgaricus OLL1247 (Accession No.: NITE BP-01814), Lactobacillus delbrueckii subsp. bulgaricus OLL205013 (Accession No.: NITE BP-02411), Lactobacillus delbrueckii OLL204989 (Accession No.: NITE BP-02874), Lactobacillus delbrueckii subsp. delbrueckii (JCM1012T), Lactobacillus delbrueckii subsp. indica (JCM 15610T), Lactobacillus delbrueckii subsp. lactis (JCM 1248T) and Lactobacillus delbrueckii subsp. mulberry (JCM 1248T). 17838T) in the group consisting of any one or more.

7. The use according to any one of claims 1 to 3, wherein The bacteria belonging to Lactobacillus delbrueckii are living bacteria.

8. The use according to any one of claims 1 to 3, wherein The composition is fermented milk that does not contain bacteria belonging to Lactobacillus paragaseri.

9. The use according to any one of claims 1 to 3, wherein Bacteria belonging to Lactobacillus delbrueckii do not have a DNA consisting of the sequence of SEQ ID NO:

6.

10. Non-therapeutic use of bacteria belonging to Lactobacillus delbrueckii for promoting ghrelin secretion.

11. A non-therapeutic use of a bacterium belonging to Lactobacillus delbrueckii for any one or more of the following selected from the group consisting of promoting growth hormone secretion, inhibiting the function of brown adipose tissue, promoting digestive tract motility, increasing gastric acid secretion, increasing pancreatic exocrine secretion, dilating blood vessels, increasing cardiac contractility, and protecting myocardial cells.

12. The use according to claim 11, wherein Any one or more selected from the group consisting of promoting growth hormone secretion, inhibiting the function of brown adipose tissue, promoting digestive tract motility, hypersecretion of gastric acid, hypersecretion of pancreatic exocrine, dilating blood vessels, increasing cardiac contractility and protecting myocardial cells is based on promoting the secretion of gastrin.

13. The use according to any one of claims 10 to 12, wherein The bacteria belonging to Lactobacillus delbrueckii are bacteria belonging to any one of Lactobacillus delbrueckii ssp. bulgaricus, Lactobacillus delbrueckii ssp. delbrueckii, Lactobacillus delbrueckii ssp. indicus, Lactobacillus delbrueckii ssp. lactis, and Lactobacillus delbrueckii ssp. sunkii.

14. The use according to any one of claims 10 to 12, wherein The bacteria belonging to Lactobacillus delbrueckii have a 16S rRNA gene having 90% or more identity with the base sequence represented by any one of SEQ ID NOs: 1 to 5.

15. The use according to any one of claims 10 to 12, wherein The bacteria belonging to Lactobacillus delbrueckii subsp. bulgaricus are selected from Lactobacillus delbrueckii subsp. bulgaricus 2038, Lactobacillus delbrueckii subsp. bulgaricus OLL1255 (Accession No.: NITE BP-76), Lactobacillus delbrueckii subsp. bulgaricus OLL1171 (Accession No.: NITE BP-01569), Lactobacillus delbrueckii subsp. bulgaricus OLL1247 (Accession No.: NITE BP-01814), Lactobacillus delbrueckii subsp. bulgaricus OLL205013 (Accession No.: NITE BP-02411), Lactobacillus delbrueckii OLL204989 (Accession No.: NITE BP-02874), Lactobacillus delbrueckii subsp. delbrueckii (JCM 1012T), Lactobacillus delbrueckii subsp. indica (JCM 15610T), Lactobacillus delbrueckii subsp. lactis (JCM 1248T) and Lactobacillus delbrueckii subsp. mulberry (JCM 1248T). 17838T) in the group consisting of any one or more.

16. The use according to any one of claims 10 to 12, wherein The bacteria belonging to Lactobacillus delbrueckii are living bacteria.

17. The use according to any one of claims 10 to 12, wherein The composition contains bacteria belonging to the group consisting of Lactobacillus delbrueckii.

18. The use according to claim 17, wherein The composition is fermented milk that does not contain bacteria belonging to Lactobacillus paragaseri.

19. The use according to any one of claims 10 to 12, wherein Bacteria belonging to Lactobacillus delbrueckii do not have a DNA consisting of the sequence of SEQ ID NO:6.

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