Novel Bifidobacterium bifidum HEM972 strain and composition for improving intestinal environment containing said strain or a culture thereof

Abstract

The present application relates to Bifidobacterium bifidum HEM972 strain (KCTC14144BP), a composition for improving intestinal environment containing the same, and a composition for treating or preventing intestinal diseases. Bifidobacterium bifidum HEM972 strain (KCTC14144BP) according to an embodiment of the present application can improve intestinal environment and treat or prevent intestinal diseases by promoting production of beneficial short-chain fatty acids such as butyric acid and propionic acid and suppressing production of harmful short-chain fatty acids such as isobutyric acid, and can be applied to pharmaceutical compositions, food compositions, health functional food compositions, feed compositions, etc.

Description

[Technical Field]

[0001] The present application relates to Bifidobacterium bifidum HEM972 strain (KCTC14144BP), a composition containing the same for improving the intestinal environment, and a composition for treating or preventing intestinal diseases. [Background technology]

[0002] Modern people's undesirable diets and unhealthy lifestyles, such as a meat-centered diet, a diet centered on instant and processed foods, excessive intake of spicy foods, and hasty changes in food and drink intake patterns due to busy living environments, as well as harmful bacteria becoming more prevalent than beneficial bacteria as we age, disrupting the balance of intestinal bacteria and significantly affecting intestinal health, potentially leading to indigestion, constipation, and colon cancer. Therefore, various methods, such as exercise therapy and the intake of health functional foods, are being researched to prevent and improve these diseases.

[0003] The intestinal microbiota is a microbial flora, a community of microorganisms in the environment. The intestinal microbiota is known to play an important role in maintaining homeostasis in the host, such as the human immune system and metabolic substances. Chemical signals are exchanged between the intestinal microbiota and the host, and the expression of immune cells, production of neurotransmitters, and short-chain fatty acids (SCFAs) produced by the intestinal microbiota have a significant impact on the host's internal systems. In particular, probiotics / prebiotics can balance an imbalanced intestinal microbiota in the host, and the resulting metabolic products of a healthy intestinal microbiota can promote the host's health.

[0004] Research into compositions for improving the intestinal environment has resulted in the development of a composition containing an aqueous extract of red koji as an active ingredient (Korea Patent Registration No. 10-1850308) and a composition containing an ethanol extract of ginseng as an active ingredient (Korea Patent Registration No. 10-1826673). However, there is still a need for research and development into compositions that are effective in improving the intestinal environment and treating intestinal diseases.

[0005] Therefore, the inventors made extensive efforts to develop an excellent composition that can improve the intestinal environment and treat or prevent intestinal diseases, and as a result, they developed a novel bacterial strain that has the ability to promote the production of beneficial short-chain fatty acids and inhibit the production of harmful short-chain fatty acids, thereby completing the present invention. Summary of the Invention [Problem to be solved by the invention]

[0006] The present application aims to provide Bifidobacterium bifidum HEM972 strain (KCTC14144BP), a composition for improving the intestinal environment containing said strain, and a composition for treating or preventing intestinal diseases.

[0007] However, the problems to be solved by the present application are not limited to those described above, and other problems not mentioned will be clearly understood by those skilled in the art from the following description. [Means for solving the problem]

[0008] A first aspect of the present application provides Bifidobacterium bifidum strain HEM972 (KCTC14144BP).

[0009] A second aspect of the present application provides a food composition for improving intestinal environment, which comprises, as an active ingredient, Bifidobacterium bifidum HEM972 strain (KCTC14144BP) or a culture thereof.

[0010] A third aspect of the present application provides a pharmaceutical composition for treating or preventing intestinal diseases, comprising Bifidobacterium bifidum HEM972 strain (KCTC14144BP) or a culture thereof as an active ingredient. [Effects of the Invention]

[0011] Bifidobacterium bifidum HEM972 strain (KCTC14144BP) according to one embodiment of the present application can improve the intestinal environment and treat or prevent intestinal diseases by promoting the production of beneficial short-chain fatty acids such as butyric acid and propionic acid and suppressing the production of harmful short-chain fatty acids such as isobutyric acid. Therefore, the strain can be used in pharmaceutical compositions, food compositions, functional health food compositions, feed compositions, etc. [Brief explanation of the drawings]

[0012] [Figure 1] FIG. 1 is a schematic diagram showing a method for measuring the ability of novel bacterial strains to promote and reduce short-chain fatty acid production. [Figure 2] FIG. 1 shows the results of confirming the ability of novel strains to promote butyric acid production. [Figure 3] FIG. 1 shows the results of confirming the ability of novel strains to promote propionic acid production. [Figure 4] FIG. 1 shows the results of confirming the ability of novel strains to reduce isobutyric acid production. [Figure 5] FIG. 1 shows the results that the novel strain does not produce short-chain fatty acids by itself. DETAILED DESCRIPTION OF THE INVENTION

[0013] Hereinafter, embodiments of the present application will be described in detail with reference to the accompanying drawings so that those skilled in the art can easily carry out the present application. However, the present application may be embodied in various different forms and is not limited to the embodiments described herein. In the drawings, parts that are not relevant to the description are omitted for clarity, and similar parts are designated by similar reference numerals throughout the specification.

[0014] Throughout this specification, when a member is said to be "on" another member, this includes not only when the member is in contact with the other member, but also when there is another member between the two members.

[0015] Throughout the specification of this application, when a part "comprises" a certain element, this means that it may further include other elements, not excluding other elements, unless otherwise specified. Terms of degree such as "about," "substantially," and the like used throughout the specification of this application are used to mean a numerical value or a close vicinity of a numerical value when inherent manufacturing and material tolerances are presented, and are used to prevent unscrupulous infringers from unfairly taking advantage of disclosures in which precise or absolute numerical values ​​are recited to aid in the understanding of this application. Terms of degree such as "steps for" or "steps of" used throughout the specification of this application do not mean "steps for."

[0016] Throughout this specification, the term "combinations thereof" contained in a Markush phrase means a mixture or combination of one or more selected from the group of elements set forth in the Markush phrase, and is meant to include one or more selected from the group of elements.

[0017] Throughout the specification of this application, the phrase "A and / or B" means "A or B, or A and B."

[0018] Hereinafter, embodiments and examples of the present application will be described in detail with reference to the accompanying drawings, but the present application is not limited to these embodiments and examples and drawings.

[0019] A first aspect of the present application provides Bifidobacterium bifidum strain HEM972 (KCTC14144BP).

[0020] In one embodiment of the present application, the strain may promote the production of beneficial short-chain fatty acids, specifically, may increase the proportion of beneficial short-chain fatty acids in the total short-chain fatty acid content.

[0021] In one embodiment of the present application, the bacterial strain may promote the production of butyric acid, specifically, the bacterial strain may promote the production of butyric acid in the intestinal environment or intestinal flora.

[0022] The term "butyrate" as used throughout this specification is a beneficial metabolite produced by carbohydrate fermentation by intestinal microorganisms and used as an energy source for colon cells. It also plays a role in maintaining colon health and regulating the immune system by suppressing colon inflammation and promoting the function of tight junctions in colon cells. It is also known to inhibit colon cancer by affecting cell death and to have preventive effects against various intestinal diseases.

[0023] Therefore, it can be seen that the Bifidobacterium bifidum HEM972 strain of the present application increases the proportion of butyric acid among intestinal short-chain fatty acids by promoting the production of butyric acid in the intestinal environment or intestinal flora, thereby exhibiting effects of suppressing colon inflammation, regulating colon immunity, and suppressing colon cancer.

[0024] In one embodiment of the present application, the strain may promote the production of propionic acid, specifically, the strain may promote the production of propionic acid in the intestinal environment or intestinal flora.

[0025] The term "propionate" as used throughout this specification is a normal metabolite produced by carbohydrate fermentation by intestinal microorganisms and is known to be beneficial to intestinal health, such as having anti-inflammatory and anti-cancer effects, suppressing fat production, and helping with colon cell surface and irritable bowel syndrome.

[0026] Therefore, it has been found that the Bifidobacterium bifidum HEM972 strain of the present application increases the proportion of propionic acid among intestinal short-chain fatty acids by promoting the production of propionic acid in the intestinal environment or intestinal flora, thereby exhibiting anti-inflammatory effects, anti-cancer effects, lipogenesis inhibitory effects, colon cell surface improving effects, and therapeutic or preventive effects for irritable bowel syndrome.

[0027] In one embodiment of the present application, the strain may reduce the production of harmful short-chain fatty acids, specifically, may reduce the proportion of harmful short-chain fatty acids in the total short-chain fatty acid content.

[0028] In one embodiment of the present application, the strain may reduce the production of one or more selected from the group consisting of isobutyric acid and isovaleric acid.

[0029] In one embodiment of the present application, the strain may have reduced production of isobutyric acid.

[0030] The term "isobutyrate" used throughout the specification of this application is a harmful metabolite produced by abnormal fermentation of intestinal microorganisms. It is known to be closely related to poor intestinal health, as it is found in higher concentrations in patients with various intestinal diseases, specifically, irritable bowel syndrome, ulcerative colitis, colorectal cancer, celiac disease, Crohn's disease, low anterior resection syndrome (LARS), and leaky gut syndrome.

[0031] Therefore, it can be seen that the Bifidobacterium bifidum HEM972 strain of the present application reduces the proportion of isobutyric acid in intestinal short-chain fatty acids by suppressing the production of isobutyric acid in the intestinal environment or intestinal flora, thereby exhibiting therapeutic or preventive effects against various intestinal diseases.

[0032] In one embodiment of the present application, the bacterial strain may be one that is unable to produce short-chain fatty acids by itself, and specifically, the bacterial strain may promote the production of beneficial short-chain fatty acids in the intestinal environment, intestinal substances, and / or intestinal flora, etc.

[0033] In one embodiment of the present application, the bacterial strain may improve the intestinal environment or treat or prevent intestinal diseases. Specifically, the bacterial strain may be contained in various compositions, such as pharmaceutical compositions, food compositions, functional health food compositions, and feed compositions, for improving the intestinal environment or treating intestinal diseases.

[0034] A second aspect of the present application provides a food composition for improving the intestinal environment, comprising Bifidobacterium bifidum HEM972 strain (KCTC14144BP) or a culture thereof as an active ingredient. The content overlapping with the first aspect also applies to the food composition of the second aspect.

[0035] The term "improvement" as used throughout the specification of this application means any action that improves or changes the intestinal environment by administering the composition.

[0036] The term "intestinal environment improvement" used throughout the present specification refers to favorably changing the composition of intestinal microorganisms and their metabolites, improving the intestinal flora, and improving intestinal function and health. The improvement of the intestinal environment increases beneficial intestinal bacteria and beneficial bacterial metabolites, which can have effects such as vitamin synthesis, improved digestion and absorption, infection prevention, and immune strengthening, and reduces harmful bacteria and harmful bacterial metabolites, which can reduce intestinal putrefaction, bacterial toxins, and carcinogens. Furthermore, improving the intestinal environment can prevent or treat not only intestinal-related diseases such as diarrhea, constipation, and enteritis, but also cancer, obesity, diabetes, and brain-related diseases.

[0037] In one embodiment of the present application, the improvement of the intestinal environment may be one or more selected from the group consisting of an increase in microbial diversity of intestinal flora, an improvement of intestinal flora, an increase in beneficial intestinal bacteria, an improvement and promotion of intestinal health, a decrease in endotoxins and hydrogen sulfide derived from intestinal microorganisms, an increase in beneficial metabolites derived from intestinal flora, an increase in beneficial short-chain fatty acids, a decrease in harmful short-chain fatty acids, an increase in the types and number of beneficial bacteria, and a decrease in the types and number of harmful bacteria, but is not limited thereto.

[0038] In one embodiment of the present application, the composition may be a composition for improving intestinal flora, a composition for promoting beneficial intestinal bacteria, or a composition for improving intestinal health.

[0039] In one embodiment of the present application, the composition may promote the production of beneficial short-chain fatty acids, specifically, butyric acid and / or propionic acid.

[0040] In one embodiment of the present application, the composition may inhibit the production of harmful short-chain fatty acids, specifically, may inhibit the production of isobutyric acid and / or isovaleric acid, and more specifically, may inhibit the production of isobutyric acid.

[0041] In one embodiment of the present application, the composition may promote the production of beneficial short-chain fatty acids in the intestinal environment, intestinal substances and / or intestinal flora.

[0042] In one embodiment of the present application, the composition may be for treating or preventing intestinal disorders.

[0043] In one embodiment of the present application, the composition may contain Bifidobacterium bifidum HEM972 strain, live cells thereof, killed cells thereof, a culture medium thereof, a disruptant thereof, and / or an extract thereof.

[0044] The term "killed bacteria" used throughout this specification is the opposite of live bacteria and refers to live bacteria and metabolic products obtained by fermentation in a form that prevents the bacteria from growing by heat treatment or other means. Killed bacteria may contain cytoplasm, cell walls, antibacterial agents such as bacteriocin, polysaccharides, organic acids, etc. Products using killed bacteria have higher stability than live bacteria products, particularly excellent heat resistance and stability against external environments, and are advantageous in that they are easier to store and have a longer shelf life than existing live bacteria products. In addition, due to stricter regulations on the use of antibiotics, they have potential as an alternative. Furthermore, with only a handful of companies currently fully engaged in the production of killed bacteria products, they have great market potential and growth potential.

[0045] The term "culture medium" as used throughout the specification of this application means a substance obtained by culturing the strain of the present application in a known liquid medium or solid medium, and may be used interchangeably with "culture."

[0046] The term "food" as used throughout this specification includes all foods in the ordinary sense, such as meats, sausages, bread, chocolates, candies, snacks, confectionery, pizza, ramen and other noodles, gums, dairy products including ice cream, various soups, beverages, tea, health supplements, alcoholic beverages, vitamin complexes, functional health foods and health foods.

[0047] The term "health functional food" used throughout the specification of this application means food manufactured and processed using raw materials or ingredients that have functional properties beneficial to the human body in accordance with Act No. 6727 on Health Functional Foods, and "functionality" means obtaining beneficial effects for health purposes, such as regulating nutrients and physiological effects on the structure and function of the human body.

[0048] The food of the present application can be prepared by methods commonly used in the art, and may be prepared by adding raw materials and ingredients commonly used in the art. Furthermore, the dosage form of the food may be prepared without limitation as long as it is a dosage form recognized as a food. The food composition of the present invention may be prepared in various dosage forms, and since it is made from food ingredients, unlike general medicines, it has the advantage of being free of side effects that can occur with long-term drug use. Furthermore, because it is highly portable, the food of the present invention can be taken as a supplement to enhance the effects of improving the intestinal environment.

[0049] The term "health food" refers to food that has more active health maintenance and promotion effects than general foods, and "health supplement food" refers to food intended for health supplementation. In some cases, the terms "health functional food," "health food," and "health supplement" may be used interchangeably. Specifically, the term "health functional food" refers to a food product obtained by adding the Bifidobacterium bifidum HEM972 strain of the present application to food ingredients such as beverages, teas, spices, gum, and confectioneries, or by encapsulating, powdering, or preparing a suspension, and which provides specific health benefits when ingested. Unlike general medicines, these foods are made from food ingredients, and therefore have the advantage of being free of side effects that can occur with long-term drug use.

[0050] The food composition of the present application can be taken on a daily basis, and is therefore expected to be highly effective in improving the intestinal environment, making it highly useful.

[0051] The food composition may further contain a physiologically acceptable carrier, but the type of carrier is not particularly limited, and any carrier commonly used in the art can be used.

[0052] The food composition may also contain additional ingredients commonly used in food compositions to improve odor, taste, visual appearance, etc. For example, vitamins A, C, D, E, B1, B2, B6, B12, niacin, biotin, folate, panthotenic acid, etc. Minerals such as zinc (Zn), iron (Fe), calcium (Ca), chromium (Cr), magnesium (Mg), manganese (Mn), copper (Cu), and chromium (Cr) may also be included. Amino acids such as lysine, tryptophan, cysteine, and valine may also be included.

[0053] The food compositions may also contain food additives such as preservatives (potassium sorbate, sodium benzoate, salicylic acid, sodium dehydroacetate, etc.), disinfectants (bleaching powder, high-strength bleaching powder, sodium hypochlorite, etc.), antioxidants (butylhydroxyanisole (BHA), butylhydroxytoluene (BHT), etc.), colorants (tar dyes, etc.), color formers (sodium nitrite, sodium nitrite, etc.), bleaching agents (sodium sulfite), seasonings (MSG, monosodium glutamate, etc.), sweeteners (dulcin, cyclamate, saccharin, sodium, etc.), flavorings (vanillin, lactones, etc.), leavening agents (alum, potassium D-bitartrate, etc.), strengtheners, emulsifiers, thickeners (thickeners), coating agents, gum bases, anti-foaming agents, solvents, and improvers. The additives may be selected depending on the type of food and used in appropriate amounts.

[0054] The Bifidobacterium bifidum HEM972 strain of the present invention may be added directly or in combination with other foods or food ingredients, and may be used appropriately in a conventional manner. The amount of active ingredient to be added may be determined appropriately depending on the intended use (prevention, health, or therapeutic treatment). Generally, when producing a food or beverage, the food composition of the present invention may be added in an amount of 50 parts by weight or less, specifically 20 parts by weight or less, to the food or beverage. However, when taken over a long period of time for health and hygiene purposes, the content may be less than the above range, and since there is no safety issue, the active ingredient may be used in an amount greater than the above range.

[0055] An example of the food composition of the present invention may be a health drink composition. In this case, various flavorings or natural carbohydrates may be added as additional ingredients, as in conventional beverages. The natural carbohydrates may be monosaccharides such as glucose and fructose; disaccharides such as maltose and sucrose; polysaccharides such as dextrin and cyclodextrin; or sugar alcohols such as xylitol, sorbitol, and erythritol. Examples of sweeteners that may be used include natural sweeteners such as thaumatin and stevia extract; and synthetic sweeteners such as saccharin and aspartame. The proportion of the natural carbohydrates may generally be about 0.01 to 0.04 g, specifically about 0.02 to 0.03 g, per 100 mL of the health drink composition of the present invention.

[0056] In addition to the above, the health drink composition may contain various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid, pectic acid salts, alginic acid, alginic acid salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, or carbonation agents. Fruit pulp for producing natural fruit juice, fruit juice beverages, or vegetable beverages may also be included. These ingredients may be used independently or in combination. The proportion of these additives is not particularly important, but is typically selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the health drink composition of the present invention.

[0057] The food composition of the present application may contain the Bifidobacterium bifidum HEM972 strain of the present application in various weight percentages as long as it exhibits the effect of improving the intestinal environment. Specifically, the food composition may contain the Bifidobacterium bifidum HEM972 strain of the present application in an amount of 0.00001 to 100 weight % or 0.01 to 80 weight % relative to the total weight of the food composition, but is not limited to these.

[0058] In one embodiment of the present application, the food composition may be a functional health food composition.

[0059] A third aspect of the present application provides a pharmaceutical composition for treating or preventing intestinal diseases, comprising Bifidobacterium bifidum HEM972 strain (KCTC14144BP) or a culture thereof as an active ingredient. The overlapping content of the first and second aspects also applies to the pharmaceutical composition of the third aspect.

[0060] In one embodiment of the present application, the composition may contain Bifidobacterium bifidum HEM972 strain, live cells thereof, killed cells thereof, a culture medium thereof, a disruptant thereof, and / or an extract thereof.

[0061] The term "intestinal disease" as used throughout the specification of this application means any disease that is caused or can be caused by a deterioration of the intestinal environment, an imbalance in the intestinal bacteria (intestinal flora), a deterioration of intestinal health, intestinal damage, etc., and may include intestinal damage, stomach disease, digestive tract disease, digestive tract damage, etc.

[0062] In one embodiment of the present application, the intestinal disease may be one or more selected from the group consisting of inflammatory bowel disease, irritable bowel syndrome (IBS), ulcerative colitis (UC), colorectal cancer, celiac disease, Crohn's disease, low anterior resection syndrome (LARS), and leaky gut syndrome, but is not limited thereto as long as it is a disease caused by an intestinal abnormality.

[0063] The term "treatment" as used throughout the specification of this application means any action of ameliorating or improving the symptoms of an intestinal disease by administering a pharmaceutical composition containing the Bifidobacterium bifidum HEM972 strain of the present application as an active ingredient to an individual suffering from an intestinal disease.

[0064] In one embodiment of the present application, the composition may promote the production of beneficial short-chain fatty acids, specifically, butyric acid and / or propionic acid.

[0065] In one embodiment of the present application, the composition may inhibit the production of harmful short-chain fatty acids, specifically, may inhibit the production of isobutyric acid and / or isovaleric acid, and more specifically, may inhibit the production of isobutyric acid.

[0066] In one embodiment of the present application, the composition may promote the production of beneficial short-chain fatty acids in the intestinal environment, intestinal substances and / or intestinal flora.

[0067] In one embodiment of the present application, the composition may improve the intestinal environment.

[0068] In one embodiment of the present application, the pharmaceutical composition may be formulated into oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories, or sterile injection solutions by conventional methods, but is not limited thereto.

[0069] In one embodiment of the present application, when the pharmaceutical composition is formulated, it may be prepared using a commonly used diluent or excipient such as, but not limited to, a filler, extender, binder, wetting agent, disintegrant, or surfactant.

[0070] In one embodiment of the present application, solid formulations for oral administration include tablets, pills, powders, granules, capsules, etc., and such solid formulations may be prepared by mixing killed cells of the strain with at least one or more excipients, such as starch, calcium carbonate, sucrose, lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used, but are not limited to these.

[0071] In one embodiment of the present application, liquid formulations for oral administration include suspensions, oral liquids, emulsions, syrups, etc., and may contain various excipients such as wetting agents, sweeteners, flavoring agents, preservatives, etc. in addition to commonly used diluents such as water and liquid paraffin, but are not limited to these.

[0072] In one embodiment of the present application, formulations for parenteral administration may include, but are not limited to, sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. For example, non-aqueous solvents or suspensions may include, but are not limited to, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. For example, suppositories may include, but are not limited to, witepsol, macrogol, Tween 61, cocoa butter, lauric butter, and glycerol gelatin.

[0073] The pharmaceutical composition according to an embodiment of the present application may be a drug composition or a quasi-drug composition.

[0074] The term "quasi-drug" as used throughout the specification of this application means an article used for the purpose of diagnosing, curing, improving, mitigating, treating, or preventing a disease in humans or animals, which has a milder effect than a drug. For example, according to the Pharmaceutical Affairs Law, quasi-drugs exclude articles used for pharmaceutical purposes, and include products used to treat or prevent a disease in humans or animals, and products that have a mild effect on the human body or do not have a direct effect.

[0075] The quasi-drug composition of the present application may be prepared in a dosage form selected from the group consisting of body cleansers, disinfectants, detergents, kitchen cleaners, cleaning detergents, toothpaste, mouthwashes, wet tissues, detergents, soaps, hand washes, hair cleansers, hair softeners, humidifier fillers, masks, ointments, and filter fillers, but is not limited thereto.

[0076] In one embodiment of the present application, the pharmaceutical composition may be administered in a pharmaceutically effective amount. As used herein, the term "pharmaceutically effective amount" refers to an amount sufficient to treat or prevent a disease with a reasonable benefit / risk ratio applicable to medical treatment or prevention. The effective dose level may be determined based on factors including the severity of the disease, drug activity, the patient's age, weight, health, sex, and drug sensitivity, the administration time, route of administration, and excretion rate of the composition of the present invention used, the duration of treatment, drugs used in combination with or concomitantly with the composition of the present invention, and other factors well known in the medical field. The pharmaceutical composition of the present application may be administered alone or in combination with a known ingredient that is effective in treating intestinal diseases. Taking all of the above factors into consideration, it is important to administer an amount that provides maximum efficacy at the minimum dose without side effects.

[0077] In one embodiment of the present application, the dosage of the pharmaceutical composition may be determined by one skilled in the art, taking into consideration the purpose of use, severity of the disease, the patient's age, weight, sex, medical history, and the type of substance used as an active ingredient. For example, the pharmaceutical composition of the present application may be administered at about 0.1 ng to about 1,000 mg / kg, preferably 1 ng to about 100 mg / kg, per adult. The administration frequency of the composition of the present application is not particularly limited, but may be once a day or divided into several doses. The dosage or frequency of administration does not limit the scope of the present application in any way.

[0078] A fourth aspect of the present application provides a method for treating an intestinal disease, comprising administering a pharmaceutically effective amount of the pharmaceutical composition of the present application to an individual suffering from the intestinal disease. The content overlapping with the first to third aspects also applies to the treatment method of the fourth aspect.

[0079] The term "individual" as used throughout this specification may include, without limitation, mammals, including mice, livestock, humans, farmed fish, etc., that are affected by or at risk of developing enteric diseases.

[0080] In one embodiment of the present application, the individual may be a non-human.

[0081] In one embodiment of the present application, the pharmaceutical composition may be administered in a single or multiple doses in a pharmaceutically effective amount, and may be formulated and administered in the form of a liquid, powder, aerosol, injection, drip infusion (Ringer's solution), capsule, pill, tablet, suppository, or patch.

[0082] The pharmaceutical composition for preventing or treating intestinal diseases of the present application may be administered via any common route as long as it can reach the target tissue.

[0083] The pharmaceutical composition of the present application may be administered via routes such as, but not limited to, intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, intradermal administration, transdermal patch administration, oral administration, intranasal administration, intrapulmonary administration, and intrarectal administration, depending on the intended purpose. However, for oral administration, the composition may be administered in an unformulated form. Because the Bifidobacterium bifidum HEM972 strain may be denatured or decomposed by gastric acid, oral compositions may be administered orally in the form of an oral patch or in a form in which the active agent is coated or protected from degradation in the stomach. The composition may also be administered by any device that allows the active agent to be delivered to target cells. [Example]

[0084] The present invention will be described in more detail below with reference to the following examples. However, the following examples are merely illustrative to aid in understanding the present invention, and the contents of the present invention are not limited to the following examples.

[0085] Example 1: Confirmation of the ability of Bifidobacterium bifidum HEM972 to promote the production and reduction of short-chain fatty acids In order to confirm the ability of the novel strain of the present invention, Bifidobacterium bifidum HEM972 (Bifidobacterium bifidum HEM972, depository: Korea Biological Resource Center, accession number: KCTC14144BP, date of accession: February 21, 2020), to promote the production and reduction of short-chain fatty acids, the following experiment was carried out.

[0086] Example 1-1: In vitro culture conditions for analyzing the amount of short-chain fatty acids produced In order to confirm the amount of short-chain fatty acids produced by the strain of the present invention under in vitro conditions, the strain was cultured as follows.

[0087] First, an intestinal environment-simulating medium having the following composition was prepared, and then the medium was placed in an anaerobic chamber (Whitley A95 anaerobic workstation) for 24 hours to replace the air, thereby making the medium anaerobic.

[0088] -Intestinal environment-like medium: NaCl 60mM, NaHCO340mM, KCl 10mM, Hemin 5×10 -5 g / L, mucin 0.5%(w / v), L-cysteine ​​HCl 0.05%(w / v)

[0089] In each well of a 96-well plate, 5 × 10 7 30 μl of the bacterial strain corresponding to a CFU / ml concentration, 180 μl of intestinal environment-simulating medium, and 15 mg of human fecal sample were dispensed, while 210 μl of intestinal environment-simulating medium and 15 mg of human fecal sample were dispensed into the control wells (NC, negative control) that were not treated with bacteria. PB_B is a commonly used probiotic strain of the same genus, Bifidobacterium, used as a reference, and the strain HEM972 in the experimental group refers to Bifidobacterium bifidum HEM972.

[0090] The 96-well plates containing the experimental and control groups were then incubated for 18 hours in an anaerobic chamber (Whitley A95 anaerobic workstation) with stirring using a DLAB mx-m agitator. After incubation, the plates were centrifuged at 3800 rpm for 10 minutes, and 100 μl of the supernatant from each well was transferred to a 96-well cell culture plate. Next, 100 μl of GC extraction solution for metabolite extraction (GC extraction solution composition: DW 10 ml, NaCl 3.3 g, phosphoric acid 50 μl, 2-ethyl butyric acid 0.5 μl) and 50 μl of DW were dispensed into 96 gas chromatography (GC) vials, along with 50 μl of the supernatant from the 96-well cell culture plate. The microbial metabolites, acetic acid, propionic acid, butyric acid, and isobutyric acid, were analyzed by gas chromatography (GC) (Figure 1).

[0091] Example 1-2: Confirmation of butyrate production promoting ability The change in the proportion of butyric acid due to the treatment with the novel strain was confirmed using the gas chromatography analysis method described in Example 1-1. Specifically, the proportion of butyric acid may be determined as the ratio of butyric acid to the total amount of normal short-chain fatty acids present in a sample: acetic acid, propionic acid, butyric acid, isobutyric acid, valeric acid, and isovaleric acid.

[0092] In Figure 2, "Total SCFA" refers to the total amount of acetic acid, propionic acid, butyric acid, isobutyric acid, valeric acid, and isovaleric acid, "Butyrate" refers to the amount of butyric acid, and "Butyrate / Total SCFA" is the value obtained by dividing the amount of butyric acid by "Total SCFA," which represents the proportion of butyric acid. To easily confirm the increase or decrease in butyric acid in each experimental group treated with bacteria compared to the control group (NC) that was not treated with any bacteria, the Butyrate / Total SCFA value for NC was expressed as 100%, and the Butyrate / Total SCFA value for each experimental group (TG) was expressed as TG / NC, which was calculated by dividing the Butyrate / Total SCFA value by NC. In addition, to compare the butyrate / total SCFA values ​​of each experimental group with the control group (NC), statistical significance was confirmed for NC and HEM972 using one-way ANOVA (post-hoc test: Fisher's LSD), and the results are indicated in the bar graphs with * (P-value<0.05), ** (P-value<0.01), and *** (P-value<0.001).To compare the butyrate / total SCFA values ​​of HEM972 and the reference strain PB_B, statistical significance was confirmed using one-way ANOVA (post-hoc test: Fisher's LSD), and the results are indicated with # (P-value<0.05), ## (P-value<0.01), and ### (P-value<0.001).

[0093] The increase or decrease in butyric acid was confirmed, and it was confirmed that the proportion of butyric acid increased statistically significantly when treated with the HEM972 strain compared to the control group and PB_B (Figure 2).

[0094] Based on the above results, it is clear that the HEM972 strain of the present invention has the function of promoting the production of butyric acid in the intestinal environment (intestinal flora).

[0095] Example 1-3: Confirmation of propionic acid production promoting ability The gas chromatography analysis method used in Example 1-1 was used to confirm the change in the propionic acid content due to the treatment with the novel strain. Specifically, the propionic acid content can be determined as the ratio of propionic acid to the total amount of normal short chain fatty acids present in a sample: acetic acid, propionic acid, butyric acid, isobutyric acid, valeric acid, and isovaleric acid.

[0096] In Figure 3, "Total SCFA" refers to the total amount of acetic acid, propionic acid, butyric acid, isobutyric acid, valeric acid, and isovaleric acid, "Propionate" refers to the amount of propionic acid, and "Propionate / Total SCFA" is the value obtained by dividing the amount of propionic acid by "Total SCFA," which represents the proportion of propionic acid. To easily confirm the increase or decrease in propionic acid in each experimental group, which was treated with bacteria compared to the control group (NC) that was not treated with any bacteria, the Propionate / Total SCFA value for NC was expressed as 100%, and the Propionate / Total SCFA value for the experimental group (TG) was expressed as TG / NC, which was calculated by dividing the Propionate / Total SCFA value by NC. In addition, to compare the propionate / total SCFA values ​​of each experimental group and the control group (NC), statistical significance was confirmed for NC and HEM972 using one-way ANOVA (post-hoc test: Fisher's LSD), and the results are indicated in the bar graphs with * (P-value<0.05), ** (P-value<0.01), and *** (P-value<0.001).To compare the propionate / total SCFA values ​​of HEM972 and the reference strain PB_B, statistical significance was confirmed using one-way ANOVA (post-hoc test: Fisher's LSD), and the results are indicated with # (P-value<0.05), ## (P-value<0.01), and ### (P-value<0.001).

[0097] As a result of checking the increase or decrease in propionic acid, it was confirmed that the proportion of propionic acid increased statistically significantly when treated with the HEM972 strain compared to the control group and PB_B (Figure 3).

[0098] Based on the above results, it is clear that the HEM972 strain of the present invention has the function of promoting the production of propionic acid in the intestinal environment (intestinal flora).

[0099] Example 1-4: Confirmation of isobutyrate production reduction ability Using the gas chromatography analysis method used in Example 1-1 above, the change in the proportion of isobutyric acid due to treatment with the novel strain was confirmed.

[0100] "Isobutyrate" in Figure 4 refers to the amount of isobutyric acid. In order to easily confirm the increase or decrease in isobutyric acid in each experimental group, which is treated with bacteria, compared to the control group (NC) that was not treated with any bacteria, the isobutyrate value for NC is expressed as 100%, and the isobutyrate value for each experimental group (TG) is expressed as the TG / NC value, which is calculated by dividing the isobutyrate value by NC. In addition, to compare the isobutyrate values ​​of HEM972 and the control group (NC), statistical significance was confirmed for the NC and experimental groups using one-way ANOVA (post-hoc test: Fisher's LSD) and is indicated in the bar graphs with * (P-value<0.05), ** (P-value<0.01), and *** (P-value<0.001).To compare the isobutyrate values ​​of HEM972 and the reference strain PB_B, statistical significance was confirmed using one-way ANOVA (post-hoc test: Fisher's LSD) and is indicated with # (P-value<0.05), ## (P-value<0.01), and ### (P-value<0.001).

[0101] The increase or decrease in isobutyric acid was confirmed, and it was confirmed that the amount of isobutyric acid was statistically significantly reduced when treated with the HEM972 strain compared to the control group and PB_B (Figure 4).

[0102] Based on the above results, it is clear that the HEM972 strain of the present invention has the function of suppressing the production of isobutyric acid in the intestinal environment (intestinal flora).

[0103] Example 2: Confirmation of whether Bifidobacterium bifidum HEM972 can produce short-chain fatty acids by itself In Example 1, it was confirmed that the novel strain of the present invention, Bifidobacterium bifidum HEM972, increases the proportion of butyric acid and propionic acid and suppresses the production of isobutyric acid. However, the above experiment did not clearly demonstrate whether the strain itself produces butyric acid and propionic acid or whether the strain promotes the production of these short-chain fatty acids in the intestinal environment. To clarify this, the following experiment was conducted.

[0104] First, an intestinal environment-simulating medium having the following composition was prepared, and then the medium was placed in an anaerobic chamber (Whitley A95 anaerobic workstation) for 24 hours to replace the air, thereby making the medium anaerobic.

[0105] -Intestinal environment-like medium: NaCl 60mM, NaHCO340mM, KCl 10mM, Hemin 5×10 -5 g / L, mucin 0.5%(w / v), L-cysteine ​​HCl 0.05%(w / v)

[0106] Next, in a 96-well plate, experimental group A (Treatment group: HEM972) was treated with 5 × 10 7 30 μl of the bacterial strain corresponding to a CFU / ml concentration and 195 μl of intestinal environment-simulating medium were dispensed. Experimental group B (Treatment group: HEM972) was 7 30 μl of the bacterial strain (corresponding to a CFU / ml concentration), 180 μl of intestinal environment-simulating medium, and 15 mg of human fecal sample were dispensed. 225 μl of intestinal environment-simulating medium was dispensed into the negative control (NC) wells, which were not treated with the bacterial strain.

[0107] The 96-well plates containing the experimental samples were then incubated for 18 hours in an anaerobic chamber (Whitley A95 anaerobic workstation) with stirring using a DLAB mx-m agitator. After incubation, the plates were centrifuged at 3800 rpm for 10 minutes, and 100 μl of the supernatant from each well was transferred to a 96-well cell culture plate. Next, 100 μl of GC extraction solution for metabolite extraction (GC extraction solution composition: DW 10 ml, NaCl 3.3 g, phosphoric acid 50 μl, 2-ethyl butyric acid 0.5 μl) was added to 96 GC vials, along with 50 μl of DW and 50 μl of the supernatant from the 96-well cell culture plate. The production of microbial metabolites (acetic acid, propionic acid, butyric acid, isobutyric acid), as well as 2-ethylbutyric acid as an internal standard, was analyzed using gas chromatography.

[0108] As a result of the above analysis, it was confirmed that butyric acid and propionic acid were not produced in experimental group A, which was treated with only the HEM972 strain and no fecal samples, whereas butyric acid and propionic acid were produced in experimental group B, which was treated with both the HEM972 strain and fecal samples (Figure 5).

[0109] Based on the above experimental results, it can be seen that the HEM972 strain of the present invention does not have the ability to produce butyric acid and propionic acid by itself, but rather has the function of stimulating and regulating the intestinal flora (fecal sample) to promote the production of butyric acid and propionic acid.

[0110] The above description of the present application is for illustrative purposes only, and those skilled in the art will understand that the present application may be easily modified into other specific forms without changing the technical concept or essential features of the present application. Therefore, it should be understood that the above-described embodiments are illustrative in all respects and not limiting. For example, each component described as a single component may be implemented in a distributed form, and similarly, each component described as a distributed component may be implemented in a combined form.

[0111] The scope of the present application is indicated by the claims that follow rather than by the above detailed description, and all modifications or variations that fall within the meaning and scope of the claims and their equivalents should be interpreted as being included within the scope of the present application.

[0112] [Accession number] Depository institution name: Korea Institute of Bioscience and Biotechnology Accession number: KCTC14144BP Date of acceptance: 20200221

[0113] JPEG0007825736000001.jpg241170

Claims

1. Bifidobacterium bifidum HEM972 strain, whose deposit number is KCTC14144BP.

2. 2. The strain of claim 1, wherein the strain promotes the production of one or more selected from the group consisting of butyric acid and propionic acid in the presence of intestinal flora.

3. 2. The strain of claim 1, wherein the strain reduces the production of isobutyric acid in the presence of intestinal flora.

4. The strain of claim 1 , wherein the strain is unable to produce short-chain fatty acids by itself.

5. A composition for regulating short-chain fatty acids in intestinal flora, comprising, as an active ingredient, the Bifidobacterium bifidum HEM972 strain, having the deposit number KCTC14144BP, or a culture thereof.

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