IAP activating components

Abstract

To provide a new composition for IAP activation.SOLUTION: The present invention provides food and beverage products and feed each of which contains a culture cultivated with Lactobacillus johnsonii, Lactobacillus delbrueckii, or Streptococcus thermophilus as an active ingredient, exhibiting IAP activating action.SELECTED DRAWING: Figure 3

Description

[Technical Field]

[0001] This invention relates to lactic acid bacteria that enhance intestinal alkaline phosphatase (IAP) activity. [Background technology]

[0002] Intestinal alkaline phosphatase (IAP) is a dephosphorylating enzyme expressed on the brush margin of the small intestine. Decreased IAP activity has been reported to be associated with various diseases, including diabetes mellitus, chronic renal failure, metabolic syndrome, inflammatory bowel disease (IBD), Crohn's disease (CD), ulcerative colitis (UC), necrotizing enterocolitis (NEC), sepsis, and antibiotic-associated diarrhea (Non-patent documents 1 and 2). Therefore, activating IAP is expected to have preventive and ameliorative effects on these diseases.

[0003] Various substances have been reported to activate IAP. For example, these include hormones such as adrenocorticotropic hormone and thyroid hormone, lipids such as long-chain and medium-chain triacylglycerols, carbohydrates such as starch, cellulose, and oligosaccharides, and others such as amino acids, peptides, vitamins, and minerals (Non-Patent Literature 1, 2). Some of these components have been suggested to have an inhibitory effect on the various diseases mentioned above (Non-Patent Literature 1, 2). Since IAP-activating ability has been reported in food components, it is thought that improving the nutritional composition of one's daily diet can maintain high IAP activity and prevent various diseases.

[0004] Incidentally, lactic acid bacteria and bifidobacteria have been used in the production of fermented foods since ancient times, but recent research is revealing that they contribute to the enhancement of various health functions. The IAP activation effect of foods fermented with lactic acid bacteria and bifidobacteria has also been reported, and for example, Non-Patent Document 3 describes that yogurt and fermented milk using Lactobacillus casei have brush-edge enzyme activation effects. Furthermore, Patent Document 1 describes a method for activating IAP in pigs by administering Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus plantarum, and Streptococcus faecium (Claim 6). However, the IAP activation effects of Lactobacillus johnsonii, Lactobacillus delbrueckii, and Streptococcus thermophilus have not been examined in these documents. [Prior art documents] [Patent Documents]

[0005] [Patent Document 1] WO1993 / 000012 [Non-patent literature]

[0006] [Non-Patent Document 1] Lalles JP. Recent advances in intestinal alkaline phosphatase, inflammation, and nutrition. Nutr Rev.2019 October;77(10):710-724. [Non-Patent Document 2] Lalles JP. Intestinal alkaline phosphatase: multiple biological roles in maintenance of intestinal homeostasis and modulation by diet. Nutr Rev. 2010 Jun;68(6):323-32. [Non-Patent Document 3] Thoreux K, Balas D, Bouley C, Senegas-Balas F. Diet supplemented with yoghurt or milk fermented by Lactobacillus casei DN-114 001 stimulates growth and brush-border enzyme activities in mouse small intestine. Digestion. 1998 Jul-Aug;59(4):349-59. [Overview of the project] [Problems that the invention aims to solve]

[0007] The object of this invention is to provide a novel IAP activation composition. [Means for solving the problem]

[0008] As a result of diligent research to solve the above problems, the inventors of this invention discovered lactic acid bacteria that enhance IAP activation activity, and found that a composition with high IAP activation ability can be obtained by preparing a culture using these lactic acid bacteria.

[0009] The present invention is specifically as follows: The term "composition" as used herein includes preparations, food and beverages, and animal feed, etc., that can be ingested by animals (including humans). [Embodiment 1] An IAP activation composition containing bacterial cells or cultures belonging to the genera Lactobacillus or Streptococcus as active ingredients. [Embodiment 2] The IAP activating composition according to Embodiment 1, characterized in that the bacterium belonging to the genus Lactobacillus or Streptococcus is selected from the group consisting of Lactobacillus johnsonii, Lactobacillus delbrueckii, and Streptococcus thermophilus. [Embodiment 3] The IAP activating composition according to Embodiment 2, characterized in that the fungus belonging to the genus Lactobacillus or Streptococcus is at least one selected from the group consisting of Lactobacillus johnsonii SBT0305 (NITE P-03628), Lactobacillus johnsonii SBT0307 (NITE P-02982), Lactobacillus johnsonii SBT0309 (NITE P-03629), Lactobacillus johnsonii SBT2082 (NITE P-03632), Lactobacillus delbrueckii SBT0803 (NITE P-03630), and Streptococcus thermophilus SBT1016 (NITE P-03631). [Embodiment 4] An IAP-activating feed composition, an IAP-activating pharmaceutical composition, or an IAP-activating food composition comprising the IAP-activating composition described in any one of Embodiments 1 to 3. [Embodiment 5] Novel lactic acid bacteria Lactobacillus johnsonii SBT0305 (NITE P-03628), Lactobacillus johnsonii SBT0309 (NITE P-03629), Lactobacillus delbrueckii SBT0803 (NITE P-03630), Streptococcus thermophilus SBT1016 (NITE P-03631), and Lactobacillus johnsonii SBT2082 (NITE P-03632).

Effects of the Invention

[0010] According to the present invention, it is possible to provide a composition for activating IAP activity containing at least one lactic acid bacterium selected from the group consisting of Lactobacillus johnsonii, Lactobacillus delbrueckii, and Streptococcus thermophilus and a culture thereof.

Brief Description of the Drawings

[0011] [Figure 1] It is a graph showing the IAP activity (U / mg protein) when a lactic acid bacterium / bifidobacterium culture was added at 1 mg / ml to the human colon cancer-derived cell line Caco-2 cultured for 2 weeks and cultured for 1 week. [Figure 2] It is a graph showing the IAP activity (U / mg protein) when the lactic acid bacterium / bifidobacterium culture showing a strong effect in FIG. 1 was added at 1 or 10 mg / ml to the human colon cancer-derived cell line Caco-2 cultured for 2 weeks and cultured for 1 week. [Figure 3] It is a graph showing the IAP activity (U / mg protein) when a culture of Lactobacillus johnsonii, Lactobacillus delbrueckii, or Streptococcus thermophilus was added at 5 mg / ml to the human colon cancer-derived cell line Caco-2 cultured for 2 weeks and cultured for 1 week.

Modes for Carrying Out the Invention

[0012] (Cells of bacteria belonging to the genus Lactobacillus or Streptococcus) In the composition for activating IAP of the present invention, any lactic acid bacterium classified into the genus Lactobacillus and the genus Streptococcus and having the ability to activate IAP can be used. Here, the genus Lactobacillus includes the genus Ligilactobacillus, the genus Levilactobacillus, the genus Limosilactobacillus, the genus Lentilactobacillus, the genus Lacticaseibacillus, the genus Lactiplantibacillus, the genus Latilactobacillus, the genus Loigolactobacillus, the genus Schleiferilactobacillus, etc., which were formerly classified as the genus Lactobacillus.

[0013] Specifically, Ligilactobacillus acidipiscis, Lactobacillus acidophilus, Lactobacillus amylovorus, Levilactobacillus brevis, Lactobacillus crispatus, Lactobacillus delbrueckii subsp. delbrueckii, Lactobacillus delbrueckii subsp. lactis, Lactobacillus delbrueckii subsp. jakobsenii, and Limosilactobacillus Limosilactobacillus fermentum, Lactobacillus gasseri, Lactobacillus paragasseri, Lactobacillus helveticus, Lactobacillus johnsonii, Lentilactobacillus kefiri, Limosilactobacillus mucosae, Limosilactobacillus oris, Lentilactobacillus parabuchneri, Lacticaseibacillus paracasei, Lactiplantibacillus Plantarum (Lactiplantibacillus plantarum), Limosilactobacillus reuteri, Lacticacei bacillusLacticaseibacillus rhamnosus, Latilactobacillus sakei, Ligilactobacillus salivarius, Limosilactobacillus vaginalis, Loigolactobacillus coryniformis, Latilactobacillus curvatus, Schleiferilactobacillus harbinensis, Streptococcus thermophilus, Streptococcus oralis, Streptococcus parauberis, Streptococcus Examples include *Streptococcus salivarius*, but the list is not limited to these.

[0014] The Lactobacillus species used in this invention is preferably Lactobacillus delbrueckii and Lactobacillus johnsonii, and particularly preferably Lactobacillus johnsonii SBT0305 (NITE P-03628), Lactobacillus johnsonii SBT0307 (NITE P-02982), Lactobacillus johnsonii SBT0309 (NITE P-03629), Lactobacillus johnsonii SBT2082 (NITE P-03632), and Lactobacillus delbrueckii SBT0803 (NITE P-03630). The Streptococcus species used in this invention is preferably Streptococcus thermophilus, and particularly preferably Streptococcus thermophilus SBT1016 (NITE P-03631).

[0015] The above bacterial strains can be obtained from the Patent Microorganism Depository Center, Biotechnology Center, National Institute of Technology and Evaluation (Kisarazu City, Chiba Prefecture, Japan).

[0016] (Preparation of cultures of bacteria belonging to the genera Lactobacillus or Streptococcus) The bacteria belonging to the Lactobacillus or Streptococcus genus used in the IAP activating composition of the present invention can be cultured according to the standard methods for culturing each bacterium, and the desired amount can be prepared. An example of preparation is shown below. Lactobacillus lactic acid bacteria are cultured using MRS medium (Difco), and Streptococcus lactic acid bacteria are cultured using 1% glucose-containing M17 medium (Difco). The cultures are then passed through three generations in a medium (SM medium) containing 10% reduced skim milk (SM, Snow Brand Megmilk), 0.5% Meast Powder N (Asahi Group Foods), 1% glucose (Wako), 0.1% sodium ascorbate (Wako), and 0.04% L-cysteine ​​hydrochloride. The cultures obtained can be used as is, or they can be concentrated, dried, or freeze-dried before use.

[0017] (How to use) As described above, the compositions of the present invention can also use cultures subjected to concentration, drying, and freeze-drying as active ingredients, and can therefore be widely used as raw materials for pharmaceuticals, food and beverages, and animal feed. Bacteria can be cultured using pharmaceuticals, food and beverages, or animal feed as a culture medium, and the culture medium, culture, suspension, or other culture-containing product can be used as is as a pharmaceutical, food and beverage, or animal feed. Alternatively, cultures cultured in other culture media can be added to pharmaceuticals, food and beverages, or animal feed.

[0018] Examples of culture forms include not only cultures prepared using synthetic media such as MRS medium (Difco) and reduced skim milk medium, which are commonly used for culturing lactic acid bacteria, but also dairy products such as cheese, fermented milk, and dairy beverages containing lactic acid bacteria. However, these are not particularly limited.

[0019] In formulating the IAP activating composition of the present invention, excipients, stabilizers, flavoring agents, etc., that are permitted under the formulation regulations may be appropriately mixed, concentrated, and freeze-dried. These dried products, concentrates, and pastes may also be included. Furthermore, within the limits that do not interfere with the IAP activation effect of the culture, excipients, binders, disintegrants, lubricants, flavoring agents, deodorizing agents, suspending agents, coating agents, and other arbitrary drugs may be mixed in the formulation. Possible dosage forms include tablets, capsules, granules, powders, syrups, etc., and oral administration is preferred.

[0020] The IAP activating composition of the present invention may be incorporated into any food or beverage, or added to raw materials during the manufacturing process of food or beverages. Examples of food and beverages include, but are not limited to, cheese, fermented milk, dairy products, lactic acid bacteria beverages, lactic acid bacteria beverages, dairy products such as butter and margarine, milk beverages, fruit juices and soft drinks, egg products such as jelly, candy, pudding and mayonnaise, confectionery and bread such as butter cake, and various types of powdered milk, as well as infant foods and nutritional compositions.

[0021] The IAP activating composition of the present invention can be incorporated into animal feed. Similar to the aforementioned food and beverage products, it can be incorporated into any type of feed, or it can be added to the raw materials during the feed manufacturing process.

[0022] The subjects to whom the composition of the present invention is administered are not particularly limited, and it can be administered to humans, but it can also be administered to animals other than humans (e.g., dogs, cats, horses, or rabbits). When administered to humans, it can be administered to minors under 20 years of age, adults, or elderly people 65 years of age or older. The composition of the present invention can improve the intestinal environment when administered to healthy subjects.

[0023] (Method for evaluating IAP activity) The method described in the examples, i.e., the method using the human colorectal cancer-derived cell line Caco-2, can be used for evaluation. Specifically, it is as follows: (1) Caco-2 cells are cultured on a collagen-coated (Nitta Gelatin) 48-well plate. The culture medium is high glucose DMEM (Nacalai Tesque) containing 10% FBS (Gibco), 1% NEAA (Nacalai Tesque), and 1% penicillin-streptomycin (Gibco). Culture is carried out at 37°C under 5% CO2 conditions. (2) Place Caco-2 cells in a collagen-coated 48-well plate in a 0.5 × 10⁶ well. 5 Seeds are seeded in cells / well, the day of confluence is considered day 0 of culture, and the culture medium is changed every 2-3 days for 2 weeks. (3) After two weeks, change the culture medium to one containing 2 mM sodium butyrate or samples of various concentrations, and incubate for 7 days, changing the medium every 2-3 days. (4) After adding the sample, the cells cultured for 7 days were washed three times with physiological saline, and the cells were lysed by adding 1% Triton-X, 1 mM PMSF, and 10 mM TBS (pH 7.3) at a rate of 200 μl / well. The entire cell lysate was collected in a 1.5 ml tube and sonicated on ice (on time 15 sec, Amp 30%). The cells were centrifuged at 4,800 × g, 5 min, 4°C, and the supernatant was collected. The collected supernatant was used as a crude enzyme extract and stored at -80°C until ready for testing. (5) The IAP activity of the crude enzyme extract was determined by Pierce TM Measurement is performed using the PNPP Substrate Kit (37620, Thermo Fisher Scientific). Bacterial alkaline phosphatase (BAP, Takara), serially diluted for calibration with 1% Triton-X, 1 mM PMSF, and 10 mM TBS (pH 7.3), or a 10-fold diluted sample, is added to a 96-well plate at a rate of 10 μl / well. PNPP substrate solution is added at a rate of 90 μl / well, and after stirring with a plate shaker, the mixture is allowed to stand in the dark at room temperature for 30 minutes. After standing, the reaction is stopped by adding 50 μl / well of 2N NaOH, and the absorbance at 405 nm is measured using Varioskan Flash (Thermo Fisher Scientific). IAP activity (U / ml) is calculated using the calibration curve created from the BAP measurement results. (6) The protein concentration of the crude enzyme extract is determined by Pierce TM The measurement is performed using the BCA Protein Assay Kit (23225, Thermo Fisher Scientific). The IAP activity (U / ml) is corrected by the protein concentration (mg / ml), and the corrected value is taken as the IAP activity (U / mg protein).

[0024] (How to use) Examples of food and beverages that can be obtained by the manufacturing method of the present invention include fermented milk products, fermented soy milk products, pickles, natto, and alcoholic beverages.

[0025] When fermenting dairy-containing raw materials with multiple strains of IAP-enhancing bacteria, it is sufficient to set conditions suitable for the growth and enzymatic reactions of the bacteria used. Generally, fermentation at 10-42°C for several hours or 1-3 days is sufficient. The food obtained in this way is a fermented milk food containing an IAP-activating composition.

[0026] By using dairy-containing raw materials, fermentation can be achieved effectively, the fermentation time can be shortened, and the resulting fermented dairy products have excellent flavor and aroma. Examples of such fermented dairy products include cheese, yogurt, and soy milk.

[0027] The pharmaceutical product obtained by the method of the present invention may, for example, be the IAP activating composition obtained as is, or the dried powder may be used as the active ingredient. There are no particular restrictions on the drying method, but freeze-drying, which can suppress deterioration of the components, is preferred. These powders can be mixed with a suitable excipient such as lactose and formulated as a powder, tablet, pill, capsule, or syrup.

[0028] Furthermore, the feed obtained by the method of the present invention may, for example, be incorporated into any feed containing the obtained IAP activating composition, or it may be added to the raw materials during the manufacturing process.

[0029] Furthermore, the food and beverages obtained by the method of the present invention can also be used as functional foods, foods for specified health uses, nutritional functional foods, or beauty foods.

[0030] The present invention will be described in more detail below based on test examples, comparative examples, and embodiments, but the present invention is not limited to these embodiments. Unless otherwise specified, percentages indicate mass percentages. [Examples]

[0031] (Example 1) Each of the test bacteria described in (1) below was inoculated into MRS medium (Difco), 1% glucose-containing GAM broth (Nissui), M17 medium (Difco), or 1% glucose-containing M17 medium (Difco), respectively, and incubated statically at 30-37°C for 16-24 hours. The cultures of each strain were subcultured for three generations in a medium containing 10% reduced skim milk (SM, Snow Brand Megmilk), 0.5% Meast Powder N (Asahi Group Foods), 1% glucose (Wako), 0.1% sodium ascorbate (Wako), and 0.04% L-cysteine ​​hydrochloride (SM medium). After measuring the pH of the skim milk cultures of each strain, they were freeze-dried to prepare a powder and stored at -80°C until use in the test.

[0032] (1) Test bacteria Each test bacterium is shown in Table 1. [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5]

[0033] (Example 2) Each of the test bacteria described in (2) below was inoculated into MRS medium (Difco), 1% glucose-containing GAM broth (Nissui), M17 medium (Difco), and 1% glucose-containing M17 medium (Difco), respectively, and incubated statically at 30-37°C for 16-24 hours. The cultures of each strain were subcultured for three generations in a medium containing 10% reduced skim milk (SM, Snow Brand Megmilk), 0.5% Meast Powder N (Asahi Group Foods), 1% glucose (Wako), 0.1% sodium ascorbate (Wako), and 0.04% L-cysteine ​​hydrochloride (SM medium). After measuring the pH of the skim milk cultures of each strain, they were freeze-dried to prepare a powder and stored at -80°C until use in the test.

[0034] (2) Test bacteria The individual bacteria tested are shown in Table 2. [Table 2-1] [Table 2-2]

[0035] The bacterial strains described in (1) and (2) above can be obtained from the Patent Microorganism Depository Center, Biotechnology Center, National Institute of Technology and Evaluation (Kisarazu City, Chiba Prefecture, Japan).

[0036] [Test Example 1] Selection test of lactic acid bacteria and bifidobacteria with high IAP activation ability To select lactic acid bacteria with high IAP activation ability, the following evaluation was performed: Caco-2 cells were placed in a collagen-coated 48-well plate in a 0.5 × 10⁶ layer. 5 Seeds were seeded in cells / well, and the day of confluence was considered day 0 of culture. The culture medium was changed every 2-3 days and the cells were cultured for 2 weeks.

[0037] After two weeks, the culture medium was replaced with a medium containing 2 mM sodium butyrate or 1-10 mg / ml of (Example 1), and the cells were incubated for 7 days, changing the medium every 2-3 days.

[0038] Cells cultured for 7 days after sample addition were washed three times with physiological saline, and lysed by adding 1% Triton-X, 1 mM PMSF, and 10 mM TBS (pH 7.3) at a rate of 200 μl / well. The entire cell lysate was collected in a 1.5 ml tube and sonicated on ice (on time 15 sec, Amp 30%). The cells were centrifuged at 4,800 × g, 5 min, 4°C, and the supernatant was collected. The collected supernatant was used as a crude enzyme extract and stored at -80°C until use in testing.

[0039] The IAP activity of the crude enzyme extract was determined by Pierce TM Measurements were performed using the PNPP Substrate Kit (37620, Thermo Fisher Scientific). Bacterial alkaline phosphatase (BAP, Takara), serially diluted for calibration with 1% Triton-X, 1 mM PMSF, and 10 mM TBS (pH 7.3), or a 10-fold diluted sample, was added to a 96-well plate at a rate of 10 μl / well. PNPP substrate solution was added at a rate of 90 μl / well, mixed with a plate shaker, and then allowed to stand in the dark at room temperature for 30 minutes. After standing, the reaction was stopped by adding 50 μl / well of 2N NaOH, and the absorbance at 405 nm was measured using Varioskan Flash (Thermo Fisher Scientific). IAP activity (U / ml) was calculated using a calibration curve derived from the BAP measurement results.

[0040] The protein concentration of the crude enzyme extract is determined by Pierce TM The IAP activity was measured using the BCA Protein Assay Kit (23225, Thermo Fisher Scientific). The IAP activity (U / ml) was corrected by the protein concentration (mg / ml), and the corrected value was defined as the IAP activity (U / mg protein).

[0041] Intergroup comparisons with the group treated with the same concentration of SM were performed using Dunnett's test. The significance level was set at P=0.05. * P<0.05,** P < 0.01, *** P < 0.001).

[0042] The IAP activities of Caco-2 cells supplemented with various lactic acid bacteria and Bifidobacterium cultures at 1 mg / ml are shown in Fig. 1. The test was carried out in four batches (Expt. 1 - 4), and the top three cultures in each batch were selected as the cultures with high IAP activation ability.

[0043] The IAP activities of Caco-2 cells supplemented with the top three various lactic acid bacteria and Bifidobacterium cultures in Expt. 1 - 4 at 1 or 10 mg / ml are shown in Fig. 2. Under the addition condition of 10 mg / ml, compared with the SM addition group, in Expt. 1, the Lactobacillus delbrueckii SBT0803 culture, in Expt. 2, the Lactobacillus johnsonii SBT0307 culture, and in Expt. 3, the Streptococcus thermophilus SBT1016 culture addition groups significantly enhanced the IAP activity respectively. In Expt. 4, the Bifidobacterium mongoliense culture significantly enhanced the IAP activity compared with the SM addition group, but the activity was lower compared with the aforementioned Lactobacillus delbrueckii SBT0803 culture, Lactobacillus johnsonii SBT0307 culture, and Streptococcus thermophilus SBT1016 culture. That is, it was shown that Lactobacillus delbrueckii SBT0803, Lactobacillus johnsonii SBT0307, and Streptococcus thermophilus SBT1016 are bacteria that enhance the IAP activation ability of skim milk.

[0044] [Test Example 2] Test for exploring bacterial species with high IAP activation ability In Test Example 1, the following evaluation was performed to verify whether the effects observed in Lactobacillus delbrueckii SBT0803, Lactobacillus johnsonii SBT0307, ​​and Streptococcus thermophilus SBT1016 were strain-specific or common to all fungal species.

[0045] Caco-2 cells were placed in a collagen-coated 48-well plate in a 0.5 × 10⁶ arrangement. 5 Seeds were seeded in cells / well, and the day of confluence was considered day 0 of culture. The culture medium was changed every 2-3 days and the cells were cultured for 2 weeks.

[0046] After two weeks, the culture medium was replaced with a medium containing 2 mM sodium butyrate or 5 mg / ml of (Example 2), and the cultures were incubated for 7 days, changing the medium every 2-3 days.

[0047] Cells cultured for 7 days after sample addition were washed three times with physiological saline, and lysed by adding 1% Triton-X, 1 mM PMSF, and 10 mM TBS (pH 7.3) at a rate of 200 μl / well. The entire cell lysate was collected in a 1.5 ml tube and sonicated on ice (on time 15 sec, Amp 30%). The cells were centrifuged at 4,800 × g, 5 min, 4°C, and the supernatant was collected. The collected supernatant was used as a crude enzyme extract and stored at -80°C until use in the test.

[0048] The IAP activity of the crude enzyme extract was determined by Pierce TMMeasurements were performed using the PNPP Substrate Kit (37620, Thermo Fisher Scientific). Bacterial alkaline phosphatase (BAP, Takara), serially diluted for calibration with 1% Triton-X, 1 mM PMSF, and 10 mM TBS (pH 7.3), or a 10-fold diluted sample, was added to a 96-well plate at a rate of 10 μl / well. PNPP substrate solution was added at a rate of 90 μl / well, mixed with a plate shaker, and then allowed to stand in the dark at room temperature for 30 minutes. After standing, the reaction was stopped by adding 50 μl / well of 2N NaOH, and the absorbance at 405 nm was measured using Varioskan Flash (Thermo Fisher Scientific). IAP activity (U / ml) was calculated using a calibration curve derived from the BAP measurement results.

[0049] The protein concentration of the crude enzyme extract is determined by Pierce TM The IAP activity was measured using the BCA Protein Assay Kit (23225, Thermo Fisher Scientific). The IAP activity (U / ml) was corrected by the protein concentration (mg / ml), and the corrected value was defined as the IAP activity (U / mg protein).

[0050] Intergroup comparisons with the group treated with the same concentration of SM were performed using Dunnett's test. The significance level was set at P=0.05. * P<0.05, ** P<0.01, *** P<0.001).

[0051] Figure 3 shows the IAP activity of Caco-2 cells to which cultures of two strains of Lactobacillus delbrueckii, eleven strains of Lactobacillus johnsonii, and 22 strains of Streptococcus thermophilus were added at a concentration of 5 mg / ml. As a result, cultures of 10 out of 22 Streptococcus thermophilus strains, and cultures of all strains of Lactobacillus delbrueckii and Lactobacillus johnsonii used in the test, significantly increased IAP activity compared to the SM-added group. In other words, skim milk cultures of Lactobacillus delbrueckii, Lactobacillus johnsonii, and Streptococcus thermophilus were shown to have IAP-activating ability. [Industrial applicability]

[0052] According to the present invention, it is possible to provide an IAP activation composition containing a culture of a fungus belonging to the genus Lactobacillus or Streptococcus as an active ingredient.

[0053] [Reference to deposited biological materials] (1)Lactobacillus johnsonii SBT0305 (i) The name and address of the depositary institution that deposited the biological material. National Institute of Technology and Evaluation (NITE), Patent Microorganism Depositary Center 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, Japan (Postal Code 292-0818) The date on which the depositary in Roy received the biological material. March 23, 2020 (receipt date) The receipt number assigned by the depositary in Hai to the deposit NITE AP-03628 (2)Lactobacillus johnsonii SBT0309 (i) The name and address of the depositary institution that deposited the biological material. National Institute of Technology and Evaluation (NITE), Patent Microorganism Depositary Center 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, Japan (Postal Code 292-0818) The date on which the depositary in Roy received the biological material. March 23, 2020 (receipt date) The receipt number assigned by the depositary in Hai to the deposit NITE AP-03629 (3)Lactobacillus delbrueckii SBT0803 (i) The name and address of the depositary institution that deposited the biological material. National Institute of Technology and Evaluation (NITE), Patent Microorganism Depositary Center 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, Japan (Postal Code 292-0818) The date on which the depositary in Roy received the biological material. March 23, 2020 (receipt date) The receipt number assigned by the depositary in Hai to the deposit NITE AP-03630 (4)Streptococcus thermophilus SBT1016 (i) The name and address of the depositary institution that deposited the biological material. National Institute of Technology and Evaluation (NITE), Patent Microorganism Depositary Center 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, Japan (Postal Code 292-0818) The date on which the depositary in Roy received the biological material. March 23, 2020 (receipt date) The receipt number assigned by the depositary in Hai to the deposit NITE AP-03631 (5)Lactobacillus johnsonii SBT2082 (i) The name and address of the depositary institution that deposited the biological material. National Institute of Technology and Evaluation (NITE), Patent Microorganism Depositary Center 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, Japan (Postal Code 292-0818) The date on which the depositary in Roy received the biological material. March 23, 2020 (receipt date) The receipt number assigned by the depositary in Hai to the deposit NITE AP-03632

Claims

1. A composition for activating IAP, comprising bacterial cells and cultures belonging to the genera Lactobacillus or Streptococcus as active ingredients. The bacteria belonging to the genera Lactobacillus or Streptococcus are at least one selected from the group consisting of Lactobacillus johnsonii and Streptococcus thermophilus.

2. The IAP activating composition according to claim 1, characterized in that the bacterium belonging to the genus Lactobacillus or Streptococcus is at least one selected from the group consisting of Lactobacillus johnsonii SBT0305 (NITE P-03628), Lactobacillus johnsonii SBT0307 (NITE P-02982), Lactobacillus johnsonii SBT0309 (NITE P-03629), Lactobacillus johnsonii SBT2082 (NITE P-03632), and Streptococcus thermophilus SBT1016 (NITE P-03631).

3. A feed composition for activating IAP, a pharmaceutical composition for activating IAP, or a food composition for activating IAP, comprising the IAP activating composition described in claim 1 or 2.

4. Lactobacillus johnsonii SBT0305 (NITE P-03628), Lactobacillus johnsonii SBT0309 (NITE P-03629), Streptococcus thermophilus SBT1016 (NITE P-03631), or Lactobacillus johnsonii SBT2082 (NITE P-03632).

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