Bacterium, composition, and method for producing composition

Abstract

This bacterium is Bifidobacterium bifidum MCC2085 (NITE BP-03881). This composition contains the bacterium. The composition is a food or beverage composition. The food or beverage composition is a probiotic composition or a nutritional composition. The method for producing a composition includes a step for mixing the bacterium with a raw material other than the bacterium.

Description

Bacteria, compositions, and methods for producing the compositions

[0001] The present invention relates to a bacterium, a composition, and a method for producing the composition. This application claims priority to Japanese Patent Application Nos. 2023-141728 and 2023-141717, filed on August 31, 2023, the contents of both applications being incorporated herein by reference.

[0002] In recent years, active research has been conducted into the prevention of disease occurrence or health promotion by actively ingesting bacteria that have a positive effect on animals (so-called beneficial bacteria, also known as "probiotics") to improve the intestinal environment. For example, Patent Document 1 discloses probiotics that have an immunomodulatory effect and are useful for treating pathologies associated with changes in the immune system.

[0003] Research is also being conducted to improve the intestinal flora, including beneficial bacteria, to suppress the occurrence of diseases or promote health. Improvement of the intestinal flora, mainly through the promotion of the proliferation of beneficial bacteria, is known to have beneficial effects on human health, such as regulating the intestines and preventing and improving inflammatory bowel disease (Non-Patent Documents 1 to 3).

[0004] Special Publication No. 2013-521335

[0005] Cannarella et al., “Mixture of probiotics reduces inflammatory biomarkers and improves the oxidative / nitrosative profile in people with rheumatoid arthritis”, Nutrition (Burbank, Los Angeles County, Calif.), vol. 89, 2021Kim et al., “Probiotic Supplementation Improves Cognitive Function and Mood with Changes in Gut Microbiota in Community-Dwelling Older Adults: A Randomized, 76, No. 1, 2021Gomi et al., “Bifidobacterium bifidum YIT 10347 fermented milk exerts beneficial effects on gastrointestinal discomfort and symptoms in healthy adults: A double-blind, randomized, placebo-controlled study”, Journal of Dairy Science, vol. 101, No. 6, 2018

[0006] Thus, further research and development of probiotics is required. The present invention aims to provide a novel bacterium belonging to the genus Bifidobacterium bifidum, a composition containing said bacterium, and a method for producing said composition.

[0007] The present invention encompasses the following aspects. [1] A bacterium that is Bifidobacterium bifidum MCC2085 (NITE BP-03881). [2] A composition containing the bacterium according to [1]. [3] The composition according to [2], wherein the composition is a food or drink composition. [4] The composition according to [3], wherein the food or drink composition is a probiotic composition or a nutritional composition. [5] The composition according to [4], wherein the nutritional composition is a formula. [6] The composition according to [4] or [5], further comprising human milk oligosaccharides. [7] The composition according to [2], wherein the composition is a pharmaceutical composition. [8] A method for producing a composition, comprising the step of mixing the bacterium according to [1] with a raw material.

[0008] In another aspect, the present invention includes the following: [1] A composition comprising a bacterium, Bifidobacterium bifidum MCC2085 (NITE BP-03881), and a food raw material, wherein the bacterium is a viable cell, and the content of the bacterium in the composition is 1 x 10 ３ ~1 x 10 １２ [2] A method for producing a food or beverage composition containing the bacterium Bifidobacterium bifidum MCC2085 (NITE BP-03881), comprising a step of mixing at least one of the bacterium and a culture of the bacterium with a food raw material, wherein the mixing step is carried out so that the content of the bacterium in the food or beverage composition is 1 x 10 ３ ~1 x 10 １２ [3] Bifidobacterium bifidum MCC2085 (NITE BP-03881) is added to a food or beverage composition in an amount of 1 × 10 CFU / g. ３ ~1 x 10 １２ [4] Use of the bacterium Bifidobacterium bifidum MCC2085 (NITE BP-03881) for improving the intestinal flora. [5] An oral composition for delivery to the intestines, comprising Bifidobacterium bifidum MCC2085 (NITE BP-03881).

[0009] According to the above aspects, it is possible to provide novel Bifidobacterium bacteria that act as probiotics, compositions containing the bacteria, and methods for producing the compositions. The present invention provides novel Bifidobacterium bifidum MCC2085 (accession number NITE BP-03881, 79% survival rate when cultured for 120 minutes in the presence of artificial gastric fluid at pH 3.0), which has superior acid resistance compared to Bifidobacterium bifidum ATCC29521 (3.1% survival rate when cultured for 120 minutes in the presence of artificial gastric fluid at pH 3.0). Furthermore, the production amount of the novel Bifidobacterium bifidum MCC2085 (NITE BP-03881) was increased 3.2-fold over the control, while the production amount of ATCC29521 was increased 1.7-fold over the control. Thus, MCC2085 had superior serotonin producing ability compared to ATCC29521.

[0010] 1 is a graph showing the change in bacterial cell concentration (CFU / ml) of MCC2085 and ATCC29521 over time in Test Example 2. 2 is a graph showing the change in survival rate (%) of MCC2085 and ATCC29521 over time in Test Example 2. 3 is a graph showing the amount of serotonin produced by MCC2085 and ATCC29521.

[0011] Preferred embodiments for carrying out the present invention will be described below. Note that the embodiment described below shows an example of a typical embodiment of the present disclosure, and the scope of the present technology is not to be interpreted narrowly by this. Note that in this specification, when a numerical range is expressed as "lower limit value to upper limit value," it means that both the upper limit value and the lower limit value are included.

[0012] Bifidobacterium bacteria have been reported to have various physiological functions. These physiological functions are reportedly related to intestinal proliferation or substances produced by Bifidobacterium bacteria (e.g., acetic acid, etc.). Therefore, the Bifidobacterium bacteria of this embodiment are also highly safe and can be expected to have the efficacy commonly associated with Bifidobacterium bacteria across a wide range of age groups. For this reason, the Bifidobacterium bacteria of this embodiment can be used in a wide range of compositions, such as foods and beverages, functional foods, pharmaceuticals, and feed. Furthermore, since the Bifidobacterium bacteria of this embodiment can be expected to have probiotic effects, they can also be used for purposes such as health promotion, dietary improvement, and intestinal environment improvement (intestinal regulation).

[0013] <Bifidobacterium bifidum MCC2085 (NITE BP-03881)> Bifidobacterium bifidum MCC2085 (NITE BP-03881, hereinafter also referred to simply as "MCC2085") is a novel Bifidobacterium bifidum isolated from human infant feces as shown in Test Example 1 described below. The novel bacterium was internationally deposited as Bifidobacterium bifidum MCC2085 (accession number: NITE BP-03881) on April 13, 2023, with the National Institute of Technology and Evaluation Patent Microorganisms Depositary (NPMD) (Address: Room 122, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, Japan 292-0818). This bacterium is publicly available from the above-mentioned collection institution.

[0014] MCC2085 has a high survival rate in low pH environments, i.e., high acid resistance. Furthermore, ingestion of MCC2085 by animals is expected to have an intestinal regulating effect. As used herein, pH refers to the value measured at 25°C. As used herein, "high acid resistance" can be evaluated by calculating the survival rate in Test Example 2 described below. MCC2085 is inoculated into 1 mL of artificial gastric fluid (pH 3.0) at 1 v / v% to obtain a mixed solution. The resulting mixed solution is maintained at 37°C. The time when the container containing the mixed solution is placed in an incubator is designated as 0 minutes, and aliquots of the mixed solution are taken at 0 minutes and 120 minutes, and subjected to pour plate culture for 72 hours, followed by measurement of CFU (colony forming units) for each time point. If the ratio (%) of CFUs in the pour plate culture after 120 minutes to the CFUs in the pour plate culture at 0 minutes is 70% or more, more preferably 75% or more, it is judged that the bacteria has a "high survival rate in a low pH environment," i.e., has "high acid resistance."

[0015] MCC2085 is not limited to the bacterium itself deposited and registered with the institution under that name (hereinafter, for convenience of explanation, also referred to as the "deposited bacterium"), but also includes bacteria substantially equivalent thereto (also referred to as "derived strains" or "derived strains"). With regard to bacteria, "bacteria substantially equivalent to the deposited bacterium" means bacteria that belong to the same species as the deposited bacterium and have acid resistance equivalent to or greater than that of the deposited bacterium. Bacteria substantially equivalent to the deposited bacterium may be, for example, a derived strain of the deposited bacterium. Derivative strains include bacteria bred from the deposited bacterium and bacteria that naturally arise from the deposited bacterium.

[0016] In this embodiment, a bacterium substantially equivalent to MCC2085 refers to, for example, a bacterium of the same genus, in which the base sequence of the 16S rRNA gene has preferably 99.9% or more, more preferably 100%, homology with the base sequence of the 16S rRNA gene of MCC2085, and preferably has the same bacteriological properties as MCC2085.

[0017] In this embodiment, MCC2085 also includes mutant strains of MCC2085, as long as they have the properties of MCC2085 (e.g., high survival rate in low pH environments, high serotonin production, etc.). Furthermore, it is preferable that this mutant strain is a bacterium that has the same bacteriological properties as MCC2085 and has a survival rate in low pH environments equal to or higher than that of MCC2085. Whether a certain mutant strain has a "high survival rate in low pH environments equal to or higher than that of MCC2085" can be confirmed, for example, by the method of Test Example 2 described below.

[0018] Such mutant strains may be constructed by non-artificially introducing mutations into MCC2085. Alternatively, they may be constructed by introducing mutations into the bacterium by treatment with a mutagen such as UV, or by various genetic engineering techniques.

[0019] Substantially equivalent bacteria and derived strains include the following: (1) Bacteria determined to be the same bacteria by the Randomly Amplified Polymorphic DNA method or the Pulsed-field gel electrophoresis method (described in Probiotics in food / Health and nutritional properties and guidelines for evaluation 85, page 43) (2) Bacteria that contain only genes derived from the deposited bacteria, do not contain genes of foreign origin, and have a DNA identity of 95% or more (preferably 98% or more) (3) Bacteria that have the same characteristics as bacteria bred from the bacteria (including genetic engineering modifications, mutations, or natural mutations)

[0020] MCC2085 can be grown by culturing the bacterium. The culturing method is not particularly limited as long as it allows MCC2085 to grow, and methods commonly used for culturing Bifidobacterium bacteria can be used with appropriate modifications as necessary. For example, the culture temperature may be 30 to 50°C, preferably 35 to 45°C. The culture is preferably carried out under anaerobic conditions, for example, while aerating with anaerobic gas such as carbon dioxide. The culture may also be carried out under microaerobic conditions, such as liquid static culture.

[0021] The culture medium for growing MCC2085 is not particularly limited, and media commonly used for culturing Bifidobacterium bacteria can be used with appropriate modifications as needed. For example, sugars such as galactose, glucose, fructose, mannose, cellobiose, maltose, lactose, sucrose, trehalose, starch hydrolysates, and blackstrap molasses can be used as carbon sources. Ammonium salts such as ammonia, ammonium chloride, and ammonium nitrate, as well as nitrates, can be used as nitrogen sources. Inorganic salts such as sodium chloride, potassium chloride, potassium sulfate, magnesium sulfate, calcium chloride, calcium nitrate, manganese chloride, and ferrous sulfate can also be used. Organic components such as soybean flour, defatted soybean meal, meat extract, and yeast extract can also be used. As a prepared culture medium, for example, MRS medium can be suitably used.

[0022] After culturing MCC2085, the resulting culture can be used as is, or it can be diluted or concentrated, or the bacterial cells alone can be recovered from the culture. As long as the effects of the present invention are achieved, MCC2085 can also be used after culturing and subjected to additional treatments such as heating, drying, crushing, and sterilization. These treatments can be used in combination as appropriate. Drying methods include spray drying and freeze drying. Sterilization methods include retort sterilization, UHT (Ultra High Temperature) sterilization, autoclaving, high-pressure steam sterilization, dry heat sterilization, circulating steam disinfection, electromagnetic wave sterilization, electron beam sterilization, high-frequency sterilization, radiation sterilization, ultraviolet sterilization, ethylene oxide gas sterilization, hydrogen peroxide gas plasma sterilization, and chemical sterilization (specifically, alcohol sterilization, formalin fixation, and electrolyzed water treatment).

[0023] MCC2085 can be in the form of, for example, a bacterial cell concentrate, freeze-dried bacterial cells, spray-dried bacterial cells, sterilized bacterial cells, heat-sterilized bacterial cells, disrupted bacterial cells, or the like.

[0024] <Compositions Containing MCC2085> The compositions of this embodiment contain MCC2085. The compositions include food and beverage compositions, pharmaceutical compositions, and the like. Because MCC2085 is derived from humans, it has few side effects and is highly safe, allowing animals to ingest it continuously over a long period of time. Therefore, it can be used in products with a wide range of applications, such as food and beverage compositions and pharmaceutical compositions.

[0025] When animals ingest the composition of this embodiment, it is expected that the composition will have an effect on intestinal regulation. MCC2085 can be used by mixing live or sterilized cells with a conventional carrier or diluent that is physiologically, pharmaceutically, or food- and beverage-acceptable.

[0026] The content of MCC2085 in the entire composition of this embodiment is not particularly limited, but may be, for example, 10 ３ CFU / g or mL or more, 10 ４ CFU / g or mL or more, 10 ５ CFU / g or mL or more, or 10 ６ CFU / g or mL or more, １２ CFU / g or mL or less, 10１１ CFU / g or mL or less, or 10 １０ The content of MCC2085 in the composition of the present invention may be 10 CFU / g or mL or less. ３ CFU / g or mL ~ 10 １２ CFU / g or mL, ４ CFU / g or mL ~ 10 １１ CFU / g or mL, ５ CFU / g or mL or more to 10 １０ It may be in CFU / g or mL.

[0027] Compositions containing MCC2085 may be used for therapeutic or non-therapeutic purposes. "Non-therapeutic purposes" is a concept that does not include medical procedures, i.e., treatment of the human body through therapy. Examples include health promotion and cosmetic procedures. "Improvement" refers to the improvement of a disease, symptom, or condition; the prevention or delay of the worsening of a disease, symptom, or condition; or the reversal, prevention, or delay of the progression of a disease or symptom. "Prevention" refers to the prevention or delay of the onset of a disease or symptom in a subject, or the reduction of the risk of a disease or symptom in a subject.

[0028] When the composition of the present invention is used for non-therapeutic purposes, it can be administered to healthy subjects. A healthy subject may refer to a subject who has a healthy intestinal flora and is not suffering from a disease caused by a disturbance in the intestinal flora. Diseases caused by a disturbance in the intestinal flora include, for example, diarrhea and constipation. When the composition of the present invention is used for non-therapeutic purposes, it can be used to further improve the healthy intestinal flora of a healthy subject, and in particular, it can be used to grow MCC2085 in the intestines of a healthy subject.

[0029] When the composition of the present invention is used for non-therapeutic purposes, it is possible to regulate the intestinal flora in healthy individuals and prevent diarrhea and / or constipation.

[0030] When the composition of the present invention is used for therapeutic purposes, it can be administered to unhealthy subjects, such as those suffering from diseases caused by disturbance of the intestinal flora.

[0031] When the composition of the present invention is used for therapeutic purposes, it is possible to regulate the intestines of unhealthy people, and improve, prevent, and treat diarrhea and constipation.

[0032] When the composition of the present invention is used for therapeutic purposes, it can be used to improve the intestinal flora of unhealthy individuals, and in particular, to grow MCC2085 in the intestines of unhealthy individuals.

[0033] The composition containing MCC2085 may be used for human or non-human animals (preferably mammals), preferably humans and pets, more preferably humans. Furthermore, the target population of the present technology is not particularly limited as long as it is anyone desiring a probiotic effect, and examples include infants, children, adults, middle-aged and elderly people, the elderly, healthy individuals, and individuals with poor intestinal environments. Of these, the present technology is preferably used for infants, adults, the elderly, and individuals with poor intestinal environments.

[0034] MCC2085 is preferably administered or taken continuously for at least one week, more preferably at least four weeks, and is preferably taken daily during the administration or intake period.

[0035] The amount of MCC2085 used is not particularly limited because it is highly safe. For example, ５ ~1 x 10 １２ CFU / kg body weight / day is preferred, 1 x 10 ７ ~1 x 10 １１ CFU / kg body weight / day is more preferred, 1 x 10 ８ ~1 x 10 １０ CFU / kg body weight / day is more preferable. Alternatively, the amount used (in other words, the dosage) per individual (body weight) is 10 ７ ~10 １４ CFU / day is preferred, 10 ８ ~10 １３ CFU / day is more preferred, 10 ９ ~10 １２ CFU / day is more preferred. If MCC2085 is sterile, CFU can be substituted with cells.

[0036] The amount of MCC2085 used is not particularly limited because it is highly safe, but is preferably 0.01 to 10,000 mg / kg body weight / day, more preferably 0.01 to 1,000 mg / kg body weight / day, and even more preferably 0.1 to 100 mg / kg body weight / day, in terms of dry weight of the bacterial cells. The amount of MCC2085 used is not particularly limited because it is highly safe, but is preferably 0.001 to 100 mL / kg body weight / day, and more preferably 0.1 to 10 mL / kg body weight / day.

[0037] <Pharmaceutical Composition> The pharmaceutical composition of this embodiment is not particularly limited as long as it contains MCC2085. The pharmaceutical composition can also be used as a composition for intestinal regulation containing MCC2085.

[0038] The pharmaceutical composition of this embodiment is used for the above-mentioned therapeutic purposes in unhealthy individuals.

[0039] The dosage form of the pharmaceutical composition of this embodiment is not particularly limited. Specific examples of dosage forms of the pharmaceutical composition include tablets, pills, powders, liquids, suspensions, emulsions, granules, capsules, syrups, suppositories, injections, ointments, patches, eye drops, and nasal drops. Furthermore, additives commonly used as pharmaceutical carriers, such as excipients, binders, disintegrants, lubricants, stabilizers, flavorings, diluents, surfactants, and injection solvents, can be used in the formulation.

[0040] Furthermore, when formulating, the pharmaceutical composition according to this embodiment can contain ingredients such as excipients, pH adjusters, colorants, and flavoring agents that are commonly used in formulations. Furthermore, as long as the effects of the present invention are not impaired, the pharmaceutical composition according to this embodiment can also contain ingredients that have the effect of preventing, ameliorating, and / or treating known or future diseases or symptoms related to this embodiment. Additionally, formulation can be carried out by known methods as appropriate depending on the dosage form. When formulating, a pharmaceutical carrier may be added as appropriate.

[0041] The content of MCC2085 in the pharmaceutical composition of this embodiment is appropriately determined depending on the dosage form, dosage method, age and sex of the patient, type of disease, severity of disease, and other conditions, but is usually 1 × 10 ５ ~1 x 10 １２ CFU / g or 1 x 10 ５ ~1 x 10 １２ CFU / mL, preferably in the range of 1 x 10 ６ ~1 x 10 １２ CFU / g or 1 x 10 ６ ~1 x 10 １２ If MCC2085 is a fungicide, CFU / g or CFU / mL can be substituted with cells / g or cells / mL.

[0042] The timing of administration of the pharmaceutical composition of this embodiment is not particularly limited, and can be appropriately selected according to the treatment method for the target symptom or disease. It may also be administered prophylactically or for maintenance therapy. The dosage form is preferably determined depending on the formulation, the patient's age, sex, other conditions, or the severity of the patient's symptoms. In either case, the pharmaceutical composition of this embodiment can be administered once a day or multiple times a day, or may be administered once every few days or weeks.

[0043] <Food and drink composition> The food and drink composition of this embodiment is not particularly limited as long as it contains MCC2085, and may be a probiotic composition, a nutritional composition, or a general food and drink composition.

[0044] The dosage form of the food and beverage composition of this embodiment is not particularly limited, and may be powder, granules, paste, emulsion, oil drops, capsules, tablets, etc.

[0045] The food and beverage composition of this embodiment may be produced by adding MCC2085 to a known food or beverage, or may be produced as a new food and beverage composition by mixing MCC2085 into the ingredients of the food or beverage.

[0046] Furthermore, the food and beverage composition of this embodiment may be a probiotic composition containing a component having a probiotic effect or a component that supports the probiotic effect, as long as the effects of the present invention are not impaired. For example, the food and beverage composition of this embodiment can be prepared by combining MCC2085 with components such as carbohydrates such as human milk oligosaccharides; various proteins such as whey protein, casein protein, soy protein, and pea protein (pea protein), or mixtures or hydrolysates thereof; amino acids such as leucine, valine, isoleucine, and glutamine; vitamins such as vitamin B6 and vitamin C; creatine; citric acid; or fish oil.

[0047] The food and beverage composition of this embodiment can be used as a probiotic composition. The food and beverage composition of this embodiment can contain food and beverage ingredients in addition to MCC2085. Examples of food and beverage ingredients include sugar derivatives such as lactose, sucrose, glucose, mannitol, and sorbitol; starch derivatives such as corn starch, potato starch, α-starch, dextrin, and carboxymethyl starch; cellulose derivatives such as crystalline cellulose, hydroxypropyl cellulose, hydroxypropylmethylcellulose, carboxymethylcellulose, and calcium carboxymethylcellulose; gum arabic; dextran; pullulan; silicate derivatives such as light anhydrous silicic acid, synthetic aluminum silicate, and magnesium aluminometasilicate; phosphate derivatives such as calcium phosphate; carbonate derivatives such as calcium carbonate; and sulfate derivatives such as calcium sulfate.

[0048] The food and drink composition of this embodiment usually contains 1 x 10 MCC2085 per total dry weight. ３ CFU / g ~ 1 x 10 １２ CFU / g, more preferably 1 x 10 ５ CFU / g ~ 1 x 10 １１ It is possible to include CFU / g.

[0049] The food and beverage composition of this embodiment is not particularly limited as long as it contains MCC2085. Examples of food and beverage compositions include beverages such as soft drinks, carbonated drinks, nutritional drinks, fruit juice drinks, and lactic acid bacteria drinks (including concentrated concentrates and powders for preparation of these beverages); frozen desserts such as ice cream, sherbet, and shaved ice; sweets such as candy, candy, gum, chocolate, tablet sweets, snacks, biscuits, jelly, jam, cream, and baked goods; dairy products such as processed milk, milk drinks, fermented milk, yogurt drinks, and butter; and bread. The food and beverage composition may also be a supplement, for example, a tablet supplement. When it is a supplement, MCC2085 can be ingested without being affected by other foods in terms of daily food intake and calorie intake.

[0050] Furthermore, the food and beverage composition defined in this embodiment can be used as a probiotic composition. That is, the food and beverage composition of this embodiment can be provided or sold as a food or beverage labeled with a use (including health use) such as a probiotic. Furthermore, the food and beverage can be provided or sold with a label indicating the intended users of the food and beverage, such as "those who wish to live with Bifidobacterium bacteria," "those who wish to improve their intestinal environment," "those who wish to regulate their stomach condition," "those who wish to create a healthy intestinal environment," or "those who wish to ingest Bifidobacteria that reach the intestines alive." The act of "labeling" includes all acts for informing consumers of the intended use, and any expression that can evoke or infer the intended use falls under the "labeling" act of the present invention, regardless of the purpose of the labeling, the content of the labeling, the object and medium on which it is displayed, etc.

[0051] Furthermore, it is preferable that the "labeling" be done in an expression that allows consumers to directly recognize the intended use. Specifically, this includes acts of transferring, delivering, displaying for the purpose of transferring or delivering, or importing food and beverage products or product packaging on which the intended use is stated, displaying or distributing advertisements, price lists, or transaction documents related to the products and including the intended use, or providing information containing the above-mentioned content by electromagnetic means (specifically, the Internet, etc.).

[0052] On the other hand, the content of the labeling is preferably a labeling approved by the government, etc. (for example, a labeling approved based on various systems established by the government and made in a manner based on such approval.) Furthermore, it is preferable that such a labeling content be attached to promotional materials at the point of sale, such as packaging, containers, catalogs, pamphlets, and POP (Point of Purchase) notices, as well as other documents.

[0053] "Labeling" also includes labeling as health food, functional food, enteral nutritional food, special dietary food, health functional food, food for specified health uses, food with nutrient functions, or food with functional claims. Among these, labeling approved by the Consumer Affairs Agency in Japan, such as labeling approved under systems related to foods for specified health uses, foods with nutrient functions, or foods with functional claims, or similar systems, can be cited. Specific examples include labeling as a food for specified health uses, labeling as a conditional food for specified health uses, labeling claiming to affect the structure or function of the body, labeling claiming disease risk reduction, and labeling claiming functionality based on scientific evidence. More specifically, typical examples include labeling as a food for specified health uses (especially labeling for health uses) and similar labeling as defined in the Cabinet Office Ordinance on Permission for Labeling for Special Uses Provided in the Health Promotion Act (Cabinet Office Ordinance No. 57 of August 31, 2009).

[0054] The probiotic composition of the present embodiment can be administered to any subject, including unhealthy individuals, but when it is made into a food or beverage product labeled for a specific use (particularly for health purposes) or function, it is used for the above-mentioned non-therapeutic purposes.

[0055] The food and drink composition according to this embodiment can be used as a nutritional composition. In this embodiment, "nutritional composition" refers to a food and drink that is orally ingested primarily for the purpose of ingesting nutrition. The type of nutritional composition that can be used in this embodiment is not particularly limited, but is preferably formula, baby food, liquid food (enteral nutrition, porridge, etc.), sports drinks, etc., and more preferably formula. The subject of intake may be infants, toddlers, children, or adults, but is preferably infants and toddlers.

[0056] Formulated milk includes formulated powdered milk and formulated liquid milk. Formulated powdered milk is defined herein as "a powdered product obtained by processing or using as the main ingredient raw milk derived from cows, raw goat's milk, raw sheep's milk, pasteurized goat's milk, cow's milk, special cow's milk, or foods produced using these as raw ingredients, and adding nutrients necessary for infants." Formulated liquid milk is defined herein as "a liquid product obtained by processing or using as the main ingredient raw milk derived from cows, raw goat's milk, raw sheep's milk, pasteurized goat's milk, cow's milk, special cow's milk, or foods produced using these as raw ingredients, and adding nutrients necessary for infants."

[0057] The prepared milk of this embodiment may further contain human milk oligosaccharides. The human milk oligosaccharides are not particularly limited as long as they are oligosaccharides normally contained in human milk, but include 2'-fucosyllactose, 3-fucosyllactose, lactodifucotetraose, 2',3-difucosyllactose, lacto-N-triose II, lacto-N-tetraose, lacto-N-neotetraose, lacto-N-fucopentaose I, lacto-N-neofucopentaose, lacto-N-fucopentaose II, lacto-N-fucopentaose III, lacto-N-fucopentaose ... Preferred examples of such oligosaccharides include neutral human milk oligosaccharides such as lactose V, lacto-N-neofucopentaose V, lacto-N-difucohexaose I, lacto-N-difucohexaose II, 6'-galactosyllactose, 3'-galactosyllactose, lacto-N-hexaose, and lacto-N-neohexaose, and acidic human milk oligosaccharides such as 3'-sialyllactose, 6'-sialyllactose, 3-fucosyl-3'-sialyllactose, and disialyl-lacto-N-tetraose. Of these, 2'-fucosyllactose, 3-fucosyllactose, 3'-sialyllactose, and 6'-sialyllactose are preferred, with 2'-fucosyllactose being particularly preferred.

[0058] Formulated milk is a blend of nutritional ingredients such as various proteins, fats and oils, carbohydrates, minerals, and vitamins, and includes formulas processed into powder or liquid form. The nutritional ingredients can be any of the nutritional ingredients described above. Formulated milk also includes "powdered infant formula," "liquid infant formula," and "powdered milk for pregnant and nursing women," which are foods for special dietary uses as defined by the Health Promotion Act. Formulated milk also includes powdered infant formula for infants aged 0 to 12 months, follow-up milk for infants aged 6 to 9 months and older and young children (up to 3 years of age), powdered milk for low birth weight infants weighing less than 2500 g at birth (low birth weight infants), allergy formula used to treat infants with pathological conditions such as cow's milk allergy or lactose intolerance, powdered milk for lactose intolerance, powdered milk for inborn errors of metabolism, powdered milk for infants, nutritional powder for adults, and nutritional powder for the elderly.

[0059] The formula of this embodiment can be administered to any subject, including healthy and unhealthy individuals. When administered to healthy individuals, the formula of this embodiment can be used as infant formula, follow-on milk, formula for low birth weight babies, formula for young children, nutritional powder for adults, or nutritional powder for the elderly, and can be used for the above-mentioned non-therapeutic purposes.

[0060] When the formula of this embodiment is administered to unhealthy individuals, it can be used as a formula for allergies, lactose intolerance, or congenital metabolic disorders, and can be used for the above-mentioned therapeutic purposes.

[0061] The nutritional composition according to this embodiment can be applied to foods with health claims and foods for the sick. The health claim food system was established based on domestic and international trends and consistency with the existing system of foods for specified health uses, and covers not only ordinary foods but also foods in the form of tablets or capsules, etc., and is categorized into two types: foods for specified health uses, which are individually approved, and foods with nutrient functions, which are standardized.

[0062] <Method for Producing Composition> A method for producing the composition of this embodiment will be described below. In the method for producing the composition of this embodiment, the step of adding MCC2085 may be carried out at any stage in the process for producing the composition.

[0063] The method for producing the composition includes at least a step of mixing MCC2085 with raw materials. The MCC2085 described above can be used. The raw materials include the raw materials for the food and beverage compositions described above, components having probiotic effects, components that supplement the probiotic effects, and pharmaceutical carriers. In the case of a method for producing a food and beverage composition, the raw materials include at least the raw materials for the food and beverage compositions described above. In the case of a method for producing a food and beverage composition, the raw materials may include at least one of the components having probiotic effects, components that supplement the probiotic effects, and pharmaceutical carriers.

[0064] When mixing MCC2085 with the raw materials, the content of MCC2085 is 1 × 10 ３ ~1 x 10 １２ It is preferable to mix so that the CFU / g is 1 x 10 ５ ~1 x 10 １１ It is more preferable to mix them so that the concentration becomes CFU / g.

[0065] The method for producing the composition may further include a step of drying the mixture of MCC2085 and the raw materials. The drying method is not particularly limited as long as it does not impair the effects of this embodiment. Specific drying methods include spray drying and freeze-drying. The method for producing the composition may further include a step of sterilizing the mixture of MCC2085 and the raw materials. The sterilization method is not particularly limited as long as it does not impair the effects of this embodiment, and examples include retort sterilization, freeze-drying, UHT sterilization, autoclaving, high-pressure steam sterilization, dry heat sterilization, circulating steam disinfection, electromagnetic wave sterilization, electron beam sterilization, high-frequency sterilization, radiation sterilization, ultraviolet sterilization, ethylene oxide gas sterilization, hydrogen peroxide gas plasma sterilization, and chemical sterilization (specifically, alcohol sterilization, formalin fixation, and electrolyzed water treatment).

[0066] The method for producing the composition may further comprise at least one of the steps of mixing a prebiotic with the mixture and further mixing MCC2085 or a dairy component with the mixture.

[0067] The method for producing the composition may further include a step of mixing at least one type of probiotic bacteria such as Bifidobacterium bacteria other than MCC2085 and lactic acid bacteria.

[0068] The milk component is preferably in powder form, and more preferably mixed with MCC2085 bacterial powder.

[0069] The milk component is not particularly limited as long as it does not impair the effects of this embodiment. For example, milk components include cow's milk, buffalo milk, sheep's milk, goat's milk, horse's milk, skim milk, concentrated skim milk, skim milk powder, concentrated milk, whole milk powder, cream, butter, buttermilk, condensed milk, and milk proteins, and one or more selected from the group consisting of these can be used. Furthermore, the milk protein is not particularly limited, and examples thereof include whey, casein, and hydrolysates thereof, and one or more selected from the group consisting of these can be used. Milk-derived components are preferred.

[0070] The hydrolysate can be produced by hydrolyzing the milk component (preferably milk protein), such as whey hydrolysate or casein hydrolysate. Examples of hydrolysis include acid or alkali hydrolysis and enzymatic hydrolysis. Among these, enzymatic hydrolysis is preferred. Protease is preferred as the enzyme used for enzymatic hydrolysis. Examples of protease include protease, trypsin, chymotrypsin, plasmin, pepsin, papain, peptidase, and aminopeptidase. The pH during the hydrolysis reaction can be adjusted appropriately to the optimum pH for the enzyme used, for example, at a pH of 2 to 6. The temperature during the hydrolysis reaction is not particularly limited, and is typically preferably in the range of 30 to 60°C. The reaction time is preferably 2 to 10 hours.

[0071] Furthermore, when producing a supplement or tablet as the composition of the present invention, the method for producing the composition of this embodiment can include a step of mixing a culture containing MCC2085 with a pharmaceutical carrier or the like to obtain a mixture, and a step of molding the mixture into a predetermined shape for the supplement or tablet. Examples of the molding step include tableting. An example of tableting is compressing a mixture of powder or granules to obtain a tablet.

[0072] Furthermore, an example of a method for producing the composition of this embodiment is a method for producing a fermented food or drink (preferably fermented milk) containing MCC2085. A dried mixture of MCC2085 and raw materials is added to a fermented milk raw material to obtain fermented milk containing MCC2085. Alternatively, a dried mixture of MCC2085 and raw materials is mixed with fermented milk to obtain fermented milk containing MCC2085. The fermentation conditions and blending amounts are adjusted so that the bacterial cell amount of MCC2085 in the composition is the above-mentioned bacterial cell amount. This makes it possible to provide a fermented food or drink containing MCC2085, preferably fermented milk or a fermented product.

[0073] <Method for improving intestinal flora and use of bacteria for improving intestinal flora> The method for improving intestinal flora of the present invention is a method for improving intestinal flora by administering Bifidobacterium bifidum MCC2085 (NITE BP-03881) to a subject in need of oral administration at an intake of 1×10 ３ ~1 x 10 １２ The method includes administering the vaccine to a recipient so that the total number of CFU / day is 100.

[0074] In the method for improving intestinal flora of this embodiment, MCC2085 can be administered to any subject, including the above-mentioned healthy and unhealthy subjects. ３ ~1 x 10 １２ When administered to patients with intestinal problems such as constipation or diarrhea, it is expected to improve the intestinal flora. It is preferable that patients take MCC2085 continuously.

[0075] That is, the bacterium Bifidobacterium bifidum MCC2085 (NITE BP-03881) can be used to improve the intestinal flora of a subject to which it is administered.

[0076] According to another aspect of the present invention, it is possible to provide an oral composition for delivery to the intestine containing Bifidobacterium bifidum MCC2085 (NITE BP-03881). MCC2085 has high acid resistance, and therefore, when orally ingested, it can maintain a high survival rate even when coexisting with gastric acid. As a result, when MCC2085 is orally ingested, it can reach the intestine while maintaining a high survival rate. All matters related to MCC2085 can be as described above.

[0077] The present embodiment will be described in more detail below based on examples. Note that the examples described below are representative examples of the present embodiment, and should not be construed as narrowing the scope of the present invention.

[0078] [Test Example 1] (1) Objective Test Example 1 was conducted to obtain Bifidobacterium bifidum MCC2085 and to clarify the bacteriological properties of the bacterium. (2) Obtaining Bifidobacterium bifidum MCC2085 and its bacteriological properties MCC2085 was isolated from infant feces. MCC2085 was anaerobically cultured on BL agar medium (Eiken Chemical Co., Ltd., E-MG16, supplemented with 5% sterile defibrated horse blood) at 37°C for 48 hours, and the formed colonies were observed. The bacteriological properties are shown in Table 1.

[0079]

[0080] (3) Analysis of the 16S rRNA gene base sequence The 16S ribosomal RNA (16S rRNA) gene base sequence of MCC2085 was analyzed. A sample DNA solution for PCR was prepared from MCC2085 cultured in (2) above, and the 16S rRNA gene was amplified by PCR using this sample DNA solution to obtain a DNA amplification product, from which the 16S rRNA gene sequence of MCC2085 was determined.

[0081] Based on this base sequence information, a homology search was performed on the full length of the 16S rRNA gene sequence using the Basic Local Alignment Search Tool (BLAST) on the international base sequence database (Genbank) of the National Center for Biotechnology Information (NCBI). As a result, it was found to have 100% homology with Bifidobacterium bifidum ATCC 29521.

[0082] The term "100% homology with Bifidobacterium bifidum ATCC29521" should be understood to mean that both MCC2085 (accession number NITE BP-03881) and ATCC29521 share a 100% evolutionary relationship between their 16S rRNA gene sequences. Thus, both strains share a common evolutionary ancestor, as the present inventors have observed that they share 100% sequence identity in 16S rRNA. This demonstrates that the novel MCC2085 deposited under the designation Bifidobacterium bifidum MCC2085 (accession number NITE BP-03881) indeed belongs to the species Bifidobacterium bifidum and is evolutionarily related to ATCC29521. Furthermore, average nucleotide identity (ANI) analysis was performed on the whole genome sequences of ATCC29521 and MCC2085 using ANI blast in pyANI (v0.2.12). The results showed that the ANI values ​​for both were 99.00%, demonstrating that ATCC29521 and MCC2085 are distinct strains. As shown in the following Test Examples 2 and 3, MCC2085 exhibited surprisingly superior acid resistance compared to ATCC29521, and MCC2085 also exhibited significantly higher serotonin production. Therefore, it is clear that Bifidobacterium bifidum MCC2085 (NITE BP-03881) described herein belongs to the Bifidobacterium bifidum species but is also a distinct strain from ATCC29521.

[0083] [Test Example 2] (1) Objective Test Example 2 was carried out to evaluate the acid resistance of Bifidobacterium bifidum MCC2085.

[0084] (2) Bacteria The following two types of bacteria were used: Bifidobacterium bifidum MCC2085 (NITE BP-03881) Bifidobacterium bifidum (ATCC29521, Type stain)

[0085] ATCC 29521 is available from the American Type Culture Collection. ATCC is located at 10801 University Boulevard, Manassas, VA 20110, United States of America. (3) Preparation of Artificial Gastric Fluid: Sodium chloride and pepsin (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) were added to ultrapure water to a final concentration of 0.2% by mass and 0.35% by mass, respectively, and mixed with a stirrer. The resulting mixture was adjusted to pH 3.0 with hydrochloric acid and then filtered through a 0.45 μm pore filter to obtain artificial gastric fluid.

[0086] (4) Pre-culture of bacteria Two types of bacteria were anaerobically cultured in an MRS medium containing 0.05% by mass of cysteine ​​hydrochloride at 37°C for 16 hours.

[0087] (5) Acid Resistance Test Two types of bacteria were cultured in the presence of artificial gastric juice for 0 minutes, 30 minutes, and 120 minutes, and the number of bacteria was counted over time.

[0088] (5-1) Cultivation time: 0 minutes The bacteria after pre-culture were diluted 10 times with physiological saline, and then diluted 100 times with physiological saline twice to obtain a 100% ... ５ A 10-fold diluted solution containing the bacterial cells was prepared. ５ The 2-fold diluted bacterial cell-containing solution was diluted 10-fold twice, resulting in 10 ６ 10-fold diluted bacterial cell-containing solution, and ７ These three kinds of diluted bacterial cell-containing solutions were used as samples with a reaction time of 0 minutes with the artificial gastric juice.

[0089] (5-2) Incubation time: 30 minutes, 120 minutes. The bacteria after pre-incubation were diluted 10-fold with physiological saline, then 10-fold with artificial gastric juice, and reacted for 30 minutes or 120 minutes at 37°C using a heat block. After the reaction was completed, the bacteria were diluted 10-fold with Mitsuoka buffer, and further diluted 10-fold with physiological saline four times. ５ 10x dilution ６ 10-fold dilution, and ７ The Mitsuoka buffer contained 4.5 g of potassium dihydrogen phosphate, 6.0 g of disodium hydrogen phosphate, 0.5 g of L-cysteine ​​hydrochloride, 0.5 g of Tween (registered trademark) 80, and 1.0 g of agar per 1 L of ultrapure water.

[0090] (5-3) Measurement of viable cell count Three types of diluted bacterial cell-containing solutions prepared at each incubation time (0 min, 30 min, 120 min) were added to TOS medium (Yakult Pharmaceutical Co., Ltd.) at 100 μL (per dish) and cultured under anaerobic conditions. After the pour culture, the mixture was cultured at 37°C for 72 hours. After the culture, colonies were counted and the bacterial cell concentration (CFU / ml) was calculated. The bacterial cell concentration was determined by repeating the steps (5-1) to (5-3) above for each diluted bacterial cell solution (10 ５ , 10 ６ , 10 ７ The results were calculated by performing three replicates for each of the diluted bacterial cell-containing solutions (2-fold diluted bacterial cell-containing solutions) and determining the arithmetic mean value (n=3).

[0091] The survival rate of bacteria at each incubation time was calculated using the following formula: Survival rate (%) = bacterial cell concentration (CFU / ml) at any incubation time / bacterial cell concentration (CFU / ml) at 0 min incubation time × 100

[0092] (6) Results The results of the change in bacterial cell concentration over time are shown in Figure 1. ７ ATCC29521 is 10 times diluted bacterial cell-containing solution. ６ As shown in Figure 1, the bacterial cell concentration of MCC2085 was almost unchanged from the bacterial cell concentration at 0 hours of culture even after 120 minutes of culture. ９On the other hand, the bacterial cell concentration of ATCC29521 was 10 CFU / ml at 0 minutes of incubation. ９ CFU / ml, but after 120 minutes of incubation, ７ The number of CFU / ml was reduced to about 1.

[0093] The results of the time course of the survival rate of the bacterial cells are shown in Figure 2. As shown in Figure 2, the survival rate of MCC2085 after 120 minutes of culture was 79%. On the other hand, the survival rate of ATCC29521 after 30 minutes of culture was 23% and after 120 minutes of culture was 3.1%.

[0094] From the above, it was revealed that MCC2085 has high acid resistance to artificial gastric juice.

[0095] Test Example 3 (1) Objective Test Example 3 was carried out to quantify the amount of serotonin produced by Bifidobacterium bifidum MCC2085 (NITE BP-03881) and Bifidobacterium bifidum ATCC29521.

[0096] (2) Preparation of culture solution containing bacterial cells MRS medium (Difco ＴＭ Lactobacillus MRS Broth (product number 288130) was sterilized at 121°C for 15 minutes. MCC2085 or ATCC29521 was cultured in the sterilized MRS medium under anaerobic conditions at 37°C for 16 hours. The culture medium after the culture was centrifuged to collect the bacterial cells, which were then suspended in phosphate buffered saline (PBS) at a concentration of 1 x 10 ９ The concentration was adjusted to 100 cells / ml to obtain a solution containing MCC2085 or ATCC29521.

[0097] (3) Cultivation and seeding of RIN14B cells RIN14B cells (ATCC CRL-2059, enterochromaffin-like cells derived from rat pancreatic cancer) were cultured in RPMI 1640 medium (Fujifilm Wako Pure Chemical Industries, Ltd.) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin at 37°C and 5% CO. ２The cultured RIN14B cells were incubated with 0.25 w / v% trypsin / EDTA (Gibco) at 37°C for 5 minutes, diluted with RPMI 1640 medium supplemented with 10% FBS and 1% penicillin-streptomycin, and subcultured at 2.5 x 10 ５ This RIN14B cell-containing solution was seeded in a 24-well plate at 500 μL / well, and then incubated at 37° C., 5% CO ２ The cells were further cultured under these conditions for 48 hours.

[0098] (4) Quantification of Serotonin Production After culturing the RIN14B cells in (3) above, the cells were washed once with HBSS(+) solution (Fujifilm Wako Pure Chemical Industries, Ltd.). The MCC2085- or ATCC29521-containing solution prepared in (2) above was diluted 20-fold with RPMI 1640 medium supplemented with 1% penicillin-streptomycin, and added to the wells at 500 μL per well. The diluted solution was incubated at 37°C and 5% CO ２ The cells were cultured for 24 hours under the conditions described above. After 24 hours of culture, the culture supernatant was collected and the serotonin content in the culture supernatant was measured using an ELISA kit (manufactured by DLD, model number EA602 / 96). The same procedure as above was repeated except that the bacterial cell-containing solution was not added, and this was used as a control.

[0099] (5) Results The amounts of serotonin produced by MCC2085 and ATCC29521 are shown in Figure 3. For statistical analysis, intergroup comparisons were performed using a t-test, with p-value correction performed using Bonferroni. The amounts of serotonin produced by MCC2085 and ATCC29521 were significantly increased compared to the untreated control. The amount of serotonin produced by MCC2085 was significantly increased compared to that produced by ATCC29521. The ratio of serotonin produced by each bacterium to the control was 3.2 for MCC2085 and 1.7 for ATCC29521.

[0100] Other bacterial strains have also been tested (data not shown). The serotonin production effects of these bacterial strains were measured using an ELISA kit (DLD, model number EA602 / 96). The measured serotonin production levels in the supernatant were comparable to those of ATCC 29521. However, Bifidobacterium bifidum MCC2085 (NITE BP-03881) was superior to these test strains.

[0101] [Production Examples] Production examples of the composition, pharmaceutical composition, fermented milk, modified milk powder, and food and drink composition of this embodiment are shown below, but the compositions of this embodiment are not limited to these.

[0102] [Production Example 1] MCC2085 was cultured in MRS liquid medium (Difco ＴＭ The bacteria was added to 3 mL of Lactobacillus MRS Broth (product number 288130) and cultured anaerobically at 32-39°C for 5-25 hours. The culture was then concentrated and freeze-dried to obtain a freeze-dried bacterial powder (i.e., bacterial powder). The bacterial powder and whey protein concentrate (WPC) were homogeneously mixed to obtain a composition. 20 g of this composition was dissolved in 200 g of water to obtain a composition containing MCC2085.

[0103] [Production Example 2] MCC2085 was added to 3 mL of MRS liquid medium and cultured anaerobically at 32-39°C for 5-25 hours. The culture medium was then concentrated and freeze-dried to obtain a freeze-dried bacterial powder (i.e., bacterial powder). The bacterial powder and a dried powder of milk protein concentrate (MPC480, manufactured by Fonterra, protein content 80% by mass, casein protein:whey protein ratio approximately 8:2) were uniformly mixed to obtain a composition. 20 g of this composition was dissolved in 200 g of water to obtain a composition containing MCC2085.

[0104] [Production Example 3] MCC2085 was added to 3 mL of MRS liquid medium and cultured anaerobically at 32-39°C for 5-25 hours. The culture medium was then concentrated and freeze-dried to obtain a freeze-dried powder of bacteria (i.e., bacterial powder) containing Bifidobacterium bifidum MCC2085 (NITE BP-03881). The bacterial powder and crystalline cellulose were then placed in a stirring granulator and mixed. Purified water was then added to granulate the mixture, and the resulting granules were dried to obtain a granule (pharmaceutical composition) containing excipients. In this manner, a granule containing MCC2085 was obtained.

[0105] [Production Example 4] A method for producing fermented milk with added MCC2085 is described below. First, the raw milk ingredients, and optionally water and other ingredients, are mixed, and then preferably homogenized and heat sterilized. The homogenization and heat sterilization can be carried out by conventional methods. A lactic acid bacteria starter is added to the heat-sterilized sterilized milk preparation (i.e., inoculated), and the mixture is fermented at a predetermined fermentation temperature to obtain a fermented product. Curd is formed by fermentation.

[0106] As the lactic acid bacteria starter, for example, lactic acid bacteria commonly used in yogurt production, such as Lactobacillus bulgaricus, Lactococcus lactis, and Streptococcus thermophilus, can be used. When the pH reaches the target value, the formed curd is broken down by stirring and cooled to 10°C or below to obtain a fermented product. By cooling to 10°C or below, the activity of the lactic acid bacteria can be reduced, thereby suppressing acid production.

[0107] The fermented product obtained in the fermentation step is then heat-treated to obtain a heated fermented product. By moderately heating the fermented product, acid production by lactic acid bacteria in the heated fermented product can be suppressed. This prevents a decrease in pH during the subsequent production process and / or during storage of the concentrated fermented milk containing bifidobacteria. As a result, the survival rate of bifidobacteria can be improved.

[0108] Next, MCC2085 is added to the heated fermented product obtained in the heat treatment step. The amount of MCC2085 added is 1 x 10 ７ ~1 x 10 １１ CFU / mL is preferred, 1 x 10 ８ ~1 x 10 １０ CFU / mL is more preferred. If MCC2085 is a bactericidal organism, CFU / mL can be substituted with cells / mL.

[0109] MCC2085 is added to the heated fermented product, which is then concentrated. The concentration step can be carried out using any known concentration method. For example, centrifugation or membrane separation can be used. In the centrifugation method, whey is removed from the product to be concentrated (i.e., the heated fermented product to which MCC2085 has been added), thereby obtaining concentrated fermented milk with an increased solids concentration.

[0110] Fermented milk containing the MCC2085 obtained as described above can be produced.

[0111] [Production Example 5] The method for producing modified powdered milk containing MCC2085 is shown below.

[0112] 10 kg of desalted milk whey protein powder (Mirai Co., Ltd.), 6 kg of milk casein powder (Fonterra), 48 kg of lactose (Mirai Co., Ltd.), 920 g of mineral mixture (Tomita Pharmaceutical Co., Ltd.), 32 g of vitamin mixture (Tanabe Pharmaceutical Co., Ltd.), 500 g of lactulose (Morinaga Milk Industry Co., Ltd.), 500 g of raffinose (Nippon Beet Sugar Co., Ltd.), and 900 g of galactooligosaccharide liquid sugar (Yakult Pharmaceutical Co., Ltd.) were dissolved in 300 kg of warm water, further heated and dissolved at 90°C for 10 minutes, and 28 kg of modified fat (Taiyo Yushi Co., Ltd.) was added and homogenized. The mixture was then sterilized, concentrated, and spray-dried to prepare approximately 95 kg of modified milk powder. To this was added 100 kg of MCC2085 bacterial cell powder (10%) dispersed in starch. ８ ~10 １１ CFU / g, manufactured by Morinaga Milk Industry Co., Ltd.) was added to prepare approximately 95 kg of bifidobacteria-oligosaccharide formulated powdered milk. The prepared powdered milk was dissolved in water to prepare a formula with a total solids concentration of 14% (w / V), which is the standard formula concentration. When the number of bifidobacteria in the formula was 10 ６ ~10 ９CFU / 100 mL can be obtained.

[0113] By ingesting or administering the thus obtained powdered milk containing MCC2085, the intestinal environment of humans can be improved.

[0114] [Production Example 6] A method for producing an intestinal environment improving food containing MCC2085 is shown below.

[0115] MCC2085 is added to 3 mL of MRS liquid medium and cultured anaerobically at 32 to 39°C for 5 to 25 hours. The culture medium is then concentrated and freeze-dried to obtain a freeze-dried powder of the bacteria (i.e., bacterial powder), which is used to obtain the composition of the present embodiment. The bacterial powder of the present embodiment can also be added to foods containing a large amount of cellulose (e.g., vegetables or salads) and cooked, thereby obtaining the composition of the present embodiment.

[0116] Alternatively, the bacterial powder can be taken before, during, or after a meal. Since regular meals (meals with a staple food, side dish, and main dish) inevitably involve the intake of dietary fiber, taking the dried bacterial powder product of this embodiment together with meals can be expected to have an effect of improving the intestinal environment in adults.

[0117] If people who suffer from intestinal problems such as constipation or diarrhea continue to take the intestinal environment improving food of this embodiment, they can expect to see a greater improvement in their intestinal environment.

[0118] From the above, Bifidobacterium bifidum MCC2085 (NITE BP-03881) and a composition containing Bifidobacterium bifidum MCC2085 (NITE BP-03881) can promote the proliferation of Bifidobacterium bacteria in the intestines, and can be effectively used for forming a good intestinal flora and improving the intestinal environment.

Claims

1. Bifidobacterium bifidum MCC2085 (NITE BP-03881).

2. A composition comprising the bacterium described in claim 1.

3. The composition according to claim 2, wherein the composition is a food or beverage composition.

4. The composition according to claim 3, wherein the food or beverage composition is a probiotic composition or a nutritional composition.

5. The composition of claim 4, wherein the nutritional composition is a formula.

6. The composition of claim 4 or 5, further comprising human milk oligosaccharides.

7. The composition of claim 2, wherein the composition is a pharmaceutical composition.

8. A method for producing a composition, comprising the step of mixing the bacterium according to claim 1 with a raw material.

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