KR20230066931A - Faecalibacterium prausnitzii strain and uses thereof - Google Patents

Abstract

The present invention relates to a Faecalibacterium prausnitzii strain and uses thereof. The Faecalibacterium prausnitzii KBL1027 strain has the accession number of KCTC14231BP. The Faecalibacterium prausnitzii strain according to an example of the present invention is superior in promoting the production of anti-inflammatory cytokines compared to conventional strains, exhibits anti-inflammatory properties, and exhibits a therapeutic effect for inflammatory diseases, thereby being used in compositions for preventing, alleviating, and treating the inflammatory diseases.

Description

Faecalibacterium prausnitzii strain and uses thereof {Faecalibacterium prausnitzii strain and uses thereof}

The present invention relates to a strain of Picalibacterium prausnitzii and uses thereof.

Inside a healthy human body, about 100 trillion microorganisms, 10 times more than the number of cells in the human body, live symbiotically. The number of their genes is over 100 times that of humans, and the intestinal microflora is structured uniquely for each individual and each disease. The healthy intestinal commensal microflora has complex interactions with human cells and performs various functions, such as the development of the immune system and maintenance of intestinal homeostasis through the regulation of metabolism and immune response.

One example of the present invention is to provide a Faecalibacterium prausnitzii KBL1027 strain having accession number KCTC14231BP.

Another example of the present invention is a Faecalibacterium prausnitzii strain, a culture of the strain, a lysate of the strain, an extract of the strain, an extract of the culture, an extract of the lysate, and the strain It is to provide a composition containing at least one selected from the group consisting of extracellular polysaccharides produced by.

Another embodiment of the present invention is to provide a use for preventing, ameliorating, or treating inflammatory diseases of a P. prausnitzii strain having a therapeutic or preventive effect on inflammatory diseases.

Another example of the present invention is a Faecalibacterium prausnitzii strain having anti-inflammatory activity, a culture of the strain, a lysate of the strain, an extract of the strain, an extract of the culture, and a lysate of the strain It is to provide an extract, or an extracellular polysaccharide produced by the strain.

Another example of the present invention is a material having anti-inflammatory activity, for example, Faecalibacterium prausnitzii strain producing extracellular polysaccharides, a culture of the strain, a lysate of the strain, the strain It is to provide an extract of, an extract of the culture, an extract of the lysate, or a material derived from the strain (eg, an extracellular polysaccharide that induces the secretion of anti-inflammatory cytokines).

In order to achieve the above object, the present invention discovers a novel Picalibacterium prausnitzii KBL1027 strain isolated from healthy Koreans, and promotes excellent anti-inflammatory cytokine production in comparison to the existing Picalibacterium prausnitzii strain The symptom improvement effect of inflammatory diseases was confirmed.

An example of the present invention relates to Faecalibacterium prausnitzii KBL1027 strain having accession number KCTC14231BP.

Another example of the present invention is a Faecalibacterium prausnitzii strain having anti-inflammatory activity, a culture of the strain, a lysate of the strain, an extract of the strain, an extract of the culture, and a lysate of the strain It relates to an extract or an extracellular polysaccharide produced by the strain.

Another example of the present invention is a Faecalibacterium prausnitzii strain producing extracellular polysaccharide, a culture of the strain, a lysate of the strain, an extract of the strain, an extract of the culture, and the lysate extract, or extracellular polysaccharide produced by the strain. The extracellular polysaccharide may be a substance having anti-inflammatory activity.

Another example of the present invention is a Faecalibacterium prausnitzii strain, a culture of the strain, a lysate of the strain, an extract of the strain, an extract of the culture, an extract of the lysate, and the strain It relates to a composition comprising at least one selected from the group consisting of extracellular polysaccharides produced by

The composition may be a composition for preventing, improving, or treating inflammatory diseases.

The composition may be a food composition, a probiotics composition, or a prebiotics composition.

The composition may be a composition for improving intestinal microbial balance.

Hereinafter, the present invention will be described in more detail.

An example of the present invention relates to Faecalibacterium prausnitzii KBL1027 strain having accession number KCTC14231BP.

The strain may be used for anti-inflammatory purposes, for inducing anti-inflammatory cytokines, for enhancing close junctions between intestinal epithelial cells, for improving intestinal microbial balance, or for preventing, ameliorating, alleviating or treating inflammatory diseases.

The Picalibacterium prausnitzii KBL1027 strain may have a 16s rRNA sequence represented by SEQ ID NO: 4.

As a result of comparing the Average Nucleotide Identity (ANI) of the Picalibacterium prausnitzii KBL1027 strain and the Picalibacterium prausnitzii DSM17677 strain belonging to the same species as the strain in the present example, In spite of this, it showed low average base identity. This means that the Picalibacterium prausnitzii KBL1027 strain according to an example of the present invention is a novel strain having genetic characteristics distinct from conventional Picalibacterium prausnitzii microorganisms.

For example, the Picalibacterium prausnitzii KBL1027 strain has an Average Nucleotide Identity (ANI) of less than 100%, less than 99%, less than 98%, 97 % or less, 96% or less, 95% or less, 94% or less, 93% or less, 92% or less, 91% or less, 90% or less, 89% or less, 88% or less, 87% or less, 86% or less, or 85% It may be below. Even if the lower limit of the average base identity is not specified, a person skilled in the art will be able to clearly perform the Picalibacterium prausnitzii KBL1027 strain having a low average base identity with the DSM17677 strain, e.g. For example, the lower limit of the average base identity may be 5% or more, 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 60% or more, 70% or more, or 80% or more, but is limited thereto it is not going to be

As a result of comparative genomic analysis of the Picalibacterium prausnitzii KBL1027 strain in the present example, the Picalibacterium prausnitzii KBL1027 strain was specific compared to the Picalibacterium prausnitzii DSM17677 strain belonging to the same species as the strain. Many genes were present. This means that the Picalibacterium prausnitzii KBL1027 strain according to an example of the present invention is a novel strain having genetic characteristics distinct from conventional Picalibacterium prausnitzii microorganisms.

For example, the P. prausnitzi KBL1027 strain may contain genes related to cell wall and capsule functions. Genes related to the cell wall and capsule functions may be those that perform functions of exopolysaccharide biosynthesis or rhamnose containing glycans. For example, the Picalibacterium prausnitzii KBL1027 strain may contain one or more genes selected from the group consisting of the following (1) to (7):

(1) Exopolysaccharide biosynthesis glycosyltransferase EpsF (EC 2.4.1.-)

(2) Tyrosine-protein kinase EpsD (EC 2.7.10.2)

(3) Alpha-D-GlcNAc alpha-1,2-L-rhamnosyltransferase (EC 2.4.1.-) rgpA

(4) Alpha-L-Rha alpha-1,3-L-rhamnosyltransferase (EC 2.4.1.-) rgpB

(5) capsular polysaccharide biosynthesis protein

(6) Lipoprotein releasing system ATP-binding protein LolD

(7) Putative N-acetylgalactosaminyl-diphosphoundecaprenol glucuronosyltransferase TuaG

For example, the Picalibacterium prausnitzii KBL1027 strain may contain one or more genes selected from the group consisting of the following (1) to (13):

(1) Exopolysaccharide biosynthesis glycosyltransferase EpsF (EC 2.4.1.-)

(2) Tyrosine-protein kinase EpsD (EC 2.7.10.2)

(3) Alpha-D-GlcNAc alpha-1,2-L-rhamnosyltransferase (EC 2.4.1.-) rgpA

(4) Alpha-L-Rha alpha-1,3-L-rhamnosyltransferase (EC 2.4.1.-) rgpB

(5) capsular polysaccharide biosynthesis protein

(6) Lipoprotein releasing system ATP-binding protein LolD

(7) Putative N-acetylgalactosaminyl-diphosphoundecaprenol glucuronosyltransferase TuaG

(8) Predicted transcriptional regulator of N-Acetylglucosamine utilization, GntR family NagQ

(9) Membrane-bound lytic murein transglycosylase B (EC 3.2.1.-)

(10) Inosine-uridine preferring nucleoside hydrolase (EC 3.2.2.1)

(11) LysR family transcriptional regulator YnfL

(12) Cation efflux system protein CusA

(13) Cobalt-zinc-cadmium resistance protein CzcA

The Picalibacterium prausnitzii KBL1027 strain according to an example of the present invention may have anti-inflammatory properties, and specifically may have anti-inflammatory properties by inducing production of anti-inflammatory cytokines.

The Picalibacterium prausnitzii KBL1027 strain according to an example of the present invention may induce anti-inflammatory cytokine production. For example, when LPS and the strain are administered together, the amount of anti-inflammatory cytokine production (pg/mL) is more than 1 times, 1.5 times or more, 2 times or more, 2.5 times or more, 3 times compared to the control group administered with LPS alone. It may be more than 3.5 times, 4 times or more, more than 4 times, 4.1 times or more, 4.2 times or more, 4.3 times or more, 4.4 times or more, or 4.5 times or more. The anti-inflammatory cytokine may be IL-10.

The anti-inflammatory cytokine IL-10 is an immune regulatory cytokine, and is known to play an important role in inflammatory responses, tumor development, allergies, and autoimmune diseases. Specifically, IL-10 is an important immune-regulating cytokine produced by various cells, which not only suppresses inflammatory responses, but also activates various immune cells such as T cells, B cells, NK (natural killer) cells, antigen-presenting cells, and mast cells. functions to regulate the proliferation and differentiation of IL-10 also exhibits an immune activation function that removes infectious or non-infectious particles while minimizing the inflammatory response. In addition, IL-10 inhibits antigen-specific activity in most cells involved in allergic reactions, and inhibits the migration of inflammatory cells such as eosinophils in inflammatory sites, reducing the expression of IgE receptors, which are allergens, to monocytes or resinous cells. Effectively block allergen response in cells. Thus, the strain according to one embodiment of the present invention induces the production of the anti-inflammatory cytokine IL-10, and may exhibit activity to prevent, improve, or treat inflammatory diseases, allergies, or autoimmune diseases.

The Picalibacterium prausnitzii KBL1027 strain according to an example of the present invention may enhance tight junctions between intestinal epithelial cells. For example, when dextran sulfate sodium (DSS) and the strain are administered together, the expression level of genes related to tight junctions (eg, Zo-1 or Occludin ) exceeds 1-fold compared to the control group administered with DSS alone , 1.1-fold or more, 1.2-fold or more, 1.3-fold or more, 1.4-fold or more, or 1.5-fold or more, or tight junction-related genes (e.g. Zo-1 Or Occludin ) expression level may be more than 1-fold, 1.01-fold or more, 1.05-fold or more, 1.1-fold or more, 1.15-fold or more, 1.2-fold or more, 1.25-fold or more, or 1.3-fold or more.

When the tight junctions of intestinal epithelial cells are weakened, leaky gut, a type of autoimmune disease, may occur. Specifically, intestinal epithelial cells must establish a protective barrier to induce "tight junctions" of cells to prevent entry of other substances. -associated lymphoid tissue (GALT) refers to a phenomenon that triggers an immune response and causes inflammation. The strain according to one embodiment of the present invention can prevent, improve, or treat leaky gut or intestinal permeability by strengthening the tight junctions of intestinal epithelial cells, and prevent, improve, or alleviate inflammatory reactions. or can be treated.

The Picalibacterium prausnitzii KBL1027 strain according to an example of the present invention may improve the balance of microorganisms in the intestine. When an imbalance of microorganisms in the intestine occurs, abnormal interactions between the host and the microflora occur, and as a result, immune homeostasis is broken, and immune-mediated diseases such as autoimmunity, allergy, and chronic inflammatory diseases may occur. The Picalibacterium prausnitzii KBL1027 strain according to an example of the present invention can prevent, improve, alleviate or treat inflammatory reactions, allergic diseases, autoimmune diseases, etc. by improving the imbalance of the intestinal microflora in vivo. For example, the Picalibacterium prausnitzii KBL1027 strain may have one or more characteristics selected from the group consisting of the following (1) to (5):

(1) Inducing an increase in the diversity of flora in the intestines of animals;

(2) improving the dysbiosis of the animal's intestinal flora;

(3) induces an increase in the relative abundance of, or prevents a decrease in the relative abundance of, the strains of the genus Prevotella and/or the family Paraprevotellaceae in the intestine of animals;

(4) preventing an increase in the relative abundance of harmful bacteria, for example, strains having a lipopolysaccharide cell wall, for example, strains of the phylum Proteobacteria, in the intestinal flora of animals;

(5) preventing an increase in the relative abundance of strains of the genus Bacteroides in the intestinal flora of animals;

The Picalibacterium prausnitzii KBL1027 strain according to an example of the present invention may have an activity for preventing, improving or treating inflammatory diseases. For example, the Picalibacterium prausnitzii KBL1027 strain may have one or more characteristics selected from the group consisting of the following (1) and (2):

(1) Prevention of weight loss due to inflammatory diseases, for example, when dextran sulfate sodium (DSS) and the strain are administered together, weight loss rate (Weight change%) compared to the control group administered with DSS alone less than 1 times, 0.9 times or less, 0.8 times or less, 0.7 times or less, 0.6 times or less, or 0.5 times or less;

(2) Prevention of intestinal length reduction due to inflammatory diseases, for example, when dextran sulfate sodium (DSS) and the strain are administered together, the reduction rate (%) of intestinal length compared to the control group administered with DSS alone less than 1x, 0.9x or less, 0.8x or less, 0.7x or less, 0.6x or less, 0.5x or less, 0.4x or less, 0.3x or less, 0.2x or less, or 0.1x or less.

Another example of the present invention is a Faecalibacterium prausnitzii strain, a culture of the strain, a lysate of the strain, an extract of the strain, an extract of the culture, an extract of the lysate, and the strain It relates to a composition for preventing, improving, or treating inflammatory diseases, comprising at least one selected from the group consisting of extracellular polysaccharides produced by. The Faecalibacterium prausnitzii strain may be a Faecalibacterium prausnitzii KBL1027 strain having accession number KCTC14231BP.

The inflammatory disease is a disease accompanied by inflammation, and the disease accompanied by inflammation may be included without limitation, for example, immune disease, inflammatory bowel disease, inflammatory skin disease, inflammatory collagen vascular disease, atopic dermatitis, urticaria, allergic disease, Autoimmune disease, autoimmune inflammatory disease, autoimmune skin disease, autoimmune inflammatory skin disease, eczema, asthma, allergic asthma, bronchial asthma, rhinitis, allergic rhinitis, conjunctivitis, allergic conjunctivitis, food allergy, anaphylactic shock (anaphylactic shock), rheumatoid arthritis, rheumatoid fever, lupus, systemic lupus erythematosus, systemic scleroderma, psoriasis, psoriatic arthritis, asthma, Guilian-Barre syndrome, myasthenia gravis, dermatomyositis, polymyositis, polymyalgia Sclerosis, type 1 diabetes, autoimmune encephalomyelitis, polyarteritis nodosa, Hashimoto's thyroiditis, systemic sclerosis, temporal arteritis, childhood diabetes mellitus, alopecia areata, pemphigus, aphth stomatitis, autoimmune hemolytic anemia, Wegener's granulomatosis, Sjogren's syndrome, Addison's disease, Behçet's disease, edema, conjunctivitis, periodontitis, rhinitis, otitis media, chronic sinusitis, sore throat, tonsillitis, bronchitis, pneumonia, gastric ulcer, gastritis, colitis, gout, eczema, acne, contact dermatitis, seborrheic dermatitis , ankylosing myelitis, fibromyalgia, osteoarthritis, periarthritis, tendinitis, tenosynovitis, myositis, hepatitis, cystitis, nephritis, sepsis, vasculitis, and bursitis.

The immune disease is a disease caused by an abnormal immune system function, and may include an allergic disease and an autoimmune disease. The immune disease may be accompanied by inflammation, and the inflammatory disease may be caused by an immune abnormality. Therefore, in the present specification, the immune disease and the inflammatory disease may be used interchangeably.

The allergic disease is a state in which a reaction to an antigen is different from normal, and may be accompanied by inflammation. Specifically, basophils coupled with immunoglobulin E (IgE) produced by the immune system exposed to antigens and histamine secreted by mast cells may induce inflammation in surrounding tissues.

The allergic disease may be, for example, at least one selected from the group consisting of atopic dermatitis, dammargin, allergic rhinitis, allergic conjunctivitis, asthma, food allergy, and anaphylactic shock, but is not limited thereto.

The autoimmune disease is a state in which autoimmunity occurs in an organ or tissue due to an immune response to substances in the body, and may be accompanied by inflammation. The autoimmune disease may be, for example, an inflammatory skin disease. The inflammatory skin disease may be, for example, at least one selected from the group consisting of atopy, psoriasis, eczema, acne, contact dermatitis, and seborrheic dermatitis. The autoimmune disease may be a local autoimmune disease or a systemic autoimmune disease.

The autoimmune disease is rheumatoid arthritis, lupus, systemic lupus erythematosus, systemic scleroderma, atopic dermatitis, psoriasis, psoriatic arthritis, asthma, Guilian-Barre syndrome, myasthenia gravis, dermatomyositis, polymyositis, multiple sclerosis , type 1 diabetes mellitus, autoimmune encephalomyelitis, polyarteritis nodosa, hypothyroidism, hyperthyroidism, idiopathic thrombocytopenia, vitiligo, Hashimoto's thyroiditis, systemic sclerosis, temporal arteritis, childhood diabetes mellitus, alopecia areata, pemphigus, aphth stomatitis, autoimmune hemolytic anemia .

The atopic dermatitis refers to a chronic or recurrent inflammatory skin condition showing itching.

The Inflammatory Bowel Disease (IBD) is an inflammatory disease of the gastrointestinal tract, the cause of which is often unknown, and research on drug development for treating inflammatory bowel disease is being extensively conducted, and using a next-generation sequencing method, As a result of analyzing the intestinal flora of patients with inflammatory bowel disease and ulcerative colitis, an imbalance in the intestinal flora was found in the patient group compared to healthy subjects, such as a decrease in the diversity of the intestinal flora and a predominance of Proteobacteria.

The inflammatory bowel disease is one selected from the group consisting of inflammatory bowel disease, ulcerative colitis (UC), enteritis, irritable bowel syndrome (IBS), and Crohn's disease (CD). There may be more than one species.

Inflammatory disease according to an embodiment of the present invention may cause one or more symptoms or pathological phenomena selected from the group consisting of the following (1) to (10):

(1) induction of inflammatory cytokines;

(2) inhibition of anti-inflammatory cytokines;

(3) hyperactivation of autoimmunity;

(4) animal weight loss;

(5) reduced intestinal length in animals;

(6) weakening of the tight junctions of animal intestinal epithelial cells;

(7) dysbiosis of the intestinal flora of animals;

(8) Increased relative abundance of harmful bacteria, for example, strains having a lipopolysaccharide cell wall, for example, strains of the phylum Proteobacteria, in the intestinal flora of animals;

(9) Increased relative abundance of strains of the genus Bacteroides in the intestinal flora of animals;

(10) Decreased relative abundance of strains of the genus Prevotella and/or the family Paraprevotellaceae in the animal's intestinal flora.

The composition according to one embodiment of the present invention may have anti-inflammatory properties, and specifically, may have anti-inflammatory properties by inducing production of anti-inflammatory cytokines. For example, when LPS and the composition are administered together, the anti-inflammatory cytokine production (pg/mL) is more than 1 times, 1.5 times or more, 2 times or more, 2.5 times or more, 3 times compared to the control group administered with LPS alone. It may be more than 3.5 times, 4 times or more, more than 4 times, 4.1 times or more, 4.2 times or more, 4.3 times or more, 4.4 times or more, or 4.5 times or more. The anti-inflammatory cytokine may be IL-10.

The composition according to one embodiment of the present invention may have a feature of reinforcing tight junctions between intestinal epithelial cells of animals. For example, when dextran sulfate sodium (DSS) and the composition are administered together, the expression level of tight junction-related genes (eg, Zo-1 or Occludin ) is greater than 1-fold compared to the control group administered with DSS alone , 1.1-fold or more, 1.2-fold or more, 1.3-fold or more, 1.4-fold or more, or 1.5-fold or more, or tight junction-related genes (e.g. Zo-1 or Occludin ) expression level may be more than 1-fold, 1.01-fold or more, 1.05-fold or more, 1.1-fold or more, 1.15-fold or more, 1.2-fold or more, 1.25-fold or more, or 1.3-fold or more.

The composition according to one embodiment of the present invention may improve the balance of microorganisms in the intestine. For example, the composition may have one or more characteristics selected from the group consisting of the following (1) to (5):

(1) Inducing an increase in the diversity of flora in the intestines of animals;

(2) improving the dysbiosis of the animal's intestinal flora;

(3) preventing an increase in the relative abundance of harmful bacteria, for example, strains having a lipopolysaccharide cell wall, for example, strains of the phylum Proteobacteria, in the intestinal flora of animals;

(4) preventing an increase in the relative abundance of strains of the genus Bacteroides in the intestinal flora of animals;

(5) Inducing an increase in the relative abundance of, or preventing a decrease in the relative abundance of, strains of the genus Prevotella and/or the family Paraprevotellaceae in the animal's intestinal flora.

The composition according to one embodiment of the present invention may have an activity for preventing, improving or treating inflammatory diseases. For example, the composition may have one or more characteristics selected from the group consisting of the following (1) to (2):

(1) Prevents weight loss of animals due to inflammatory diseases, for example, when dextran sulfate sodium (DSS) and the composition are administered together, weight loss rate (Weight change) compared to the control group administered with DSS alone %) is less than 1x, 0.9x or less, 0.8x or less, 0.7x or less, 0.6x or less, or 0.5x or less;

(2) Preventing reduction of intestinal length in animals due to inflammatory diseases, for example, when dextran sulfate sodium (DSS) and the composition are administered together, the reduction rate of intestinal length compared to the control group administered with DSS alone ( %) is less than 1x, 0.9x or less, 0.8x or less, 0.7x or less, 0.6x or less, 0.5x or less, 0.4x or less, 0.3x or less, 0.2x or less, or 0.1x or less.

The composition may improve the microbial balance in the intestine. For example, the composition may increase the flora diversity in the animal's intestine. For example, the composition may increase the relative abundance of strains belonging to the genus Prevotella and/or Paraprevotellaceae in the intestine of animals. For example, the composition may reduce the relative abundance of strains of the genus Bacteroides and/or the phylum Proteobacteria in the intestine of an animal.

The composition may include an extracellular polysaccharide of Faecalibacterium prausnitzii strain. According to the present Example, it was confirmed that the Picalibacterium prausnitzii KBL1027 strain specifically produces extracellular polysaccharide, and thus the composition is an extracellular polysaccharide of the Picalibacterium prausnitzii KBL1027 strain having accession number KCTC14231BP It may contain.

Based on the experiment, the Picalibacterium prausnitzii KBL1027 strain according to the present invention was confirmed to have excellent efficacy in improving symptoms of colitis disease compared to other strains belonging to the same species based on the experiment, and adhesion between the anti-inflammatory cytokine IL-10 and intestinal epithelial cells The effects of enhancing the expression of Zo-1 and Occludin related to synaptic function and restoring intestinal flora were confirmed. In addition, the extracellular polysaccharide material specifically produced by the P. prausnitzii KBL1027 strain was experimentally and genomically confirmed, and the anti-inflammatory cytokine IL-10 production ability of the P. prausnitzii KBL1027 strain culture medium was confirmed. It was confirmed based on BMDM. These results mean that the KBL1027 strain of P. prausnitzi can be usefully used for the prevention, improvement, and treatment of inflammatory diseases.

Another example of the present invention is a Faecalibacterium prausnitzii strain having anti-inflammatory activity, a culture of the strain, a lysate of the strain, an extract of the strain, an extract of the culture, and a lysate of the strain It relates to an extract or an extracellular polysaccharide produced by the strain. The Faecalibacterium prausnitzii strain may be a Faecalibacterium prausnitzii KBL1027 strain having accession number KCTC14231BP.

Another example of the present invention is a material having anti-inflammatory activity, for example, a Faecalibacterium prausnitzii strain producing an extracellular polysaccharide, a culture of the strain, a lysate of the strain, and the strain It relates to an extract of, an extract of the culture, an extract of the lysate, or an extracellular polysaccharide produced by the strain. The Faecalibacterium prausnitzii strain may be a Faecalibacterium prausnitzii KBL1026 strain having accession number KCTC14230BP or a Faecalibacterium prausnitzii KBL1027 strain having accession number KCTC14231BP, preferably It may be a Picalibacterium prausnitzi KBL1027 strain having accession number KCTC14231BP.

Another example of the present invention is a Faecalibacterium prausnitzii strain, a culture of the strain, a lysate of the strain, an extract of the strain, an extract of the culture, an extract of the lysate, and the strain It relates to a food composition comprising at least one selected from the group consisting of extracellular polysaccharides produced by. The Picalibacterium prausnitzii strain is as described above. For example, the Faecalibacterium prausnitzii strain may be a Faecalibacterium prausnitzii KBL1027 strain having accession number KCTC14231BP.

Another example of the present invention is a Faecalibacterium prausnitzii strain, a culture of the strain, a lysate of the strain, an extract of the strain, an extract of the culture, an extract of the lysate, and the strain It relates to a probiotics and / or prebiotics composition containing at least one selected from the group consisting of extracellular polysaccharides produced by. The Picalibacterium prausnitzii strain is as described above. For example, the Faecalibacterium prausnitzii strain may be a Faecalibacterium prausnitzii KBL1027 strain having accession number KCTC14231BP.

Another example of the present invention is a Faecalibacterium prausnitzii strain, a culture of the strain, a lysate of the strain, an extract of the strain, an extract of the culture, an extract of the lysate, and the strain It relates to a composition for improving intestinal microbial balance, comprising at least one selected from the group consisting of extracellular polysaccharides produced by The Picalibacterium prausnitzii strain is as described above. For example, the Faecalibacterium prausnitzii strain may be a Faecalibacterium prausnitzii KBL1027 strain having accession number KCTC14231BP.

Another example of the present invention is a Faecalibacterium prausnitzii KBL1027 strain having accession number KCTC14231BP, a culture of the strain, a lysate of the strain, an extract of the strain, an extract of the culture, and the lysate It relates to a method for preventing, improving, or treating inflammatory diseases, comprising administering at least one selected from the group consisting of extracts of and extracellular polysaccharides produced by the strain. The Picalibacterium prausnitzi KBL1027 strain, the inflammatory disease, etc. are as described above.

Another example of the present invention is a Faecalibacterium prausnitzii KBL1027 strain having accession number KCTC14231BP, a culture of the strain, a lysate of the strain, an extract of the strain, an extract of the culture, and the lysate It relates to a method for improving the balance of microorganisms in the intestine, comprising the step of administering at least one selected from the group consisting of extracts of and extracellular polysaccharides produced by the strain. As a result of administering the strain according to an example of the present invention in the present Example, an increase in the diversity of the intestinal flora of animals was induced, and the dysbiosis of the intestinal flora of animals was improved. Specifically, in the intestinal flora of animals Prevention of an increase in the relative abundance of harmful bacteria, for example, strains having a lipopolysaccharide cell wall, for example, strains of the phylum Proteobacteria, and an increase in the relative abundance of strains of the genus Bacteroides in the intestinal flora of animals In addition, an increase in the relative abundance of the genus Prevotella and / or a strain of the family Paraprevotellaceae in the intestinal flora of the animal was induced, or a decrease in the relative abundance was prevented. Thus, the balance of microorganisms in the intestines of animals can be improved by administering the strain according to one embodiment of the present invention.

In the present invention, the term 'active ingredient' refers to a component that exhibits the desired activity alone or that can exhibit activity together with a carrier having no activity itself.

In the present invention, the term 'prevention' means suppressing or delaying the onset of a disease, disorder or disease. Prevention can be considered complete when the onset of a disease, disorder or condition is suppressed or delayed for a pre-determined period of time.

As used herein, the term 'treatment' refers to partially or completely alleviating, ameliorating, alleviating, inhibiting, or delaying, reducing the severity of, or reducing the severity of, a specific disease, disorder, and/or disease or symptoms of one or more symptoms or characteristics. means to reduce incidence.

The composition of the present invention, for example, a pharmaceutical composition, may further include at least one active ingredient exhibiting the same or similar function in addition to the above active ingredient.

In addition, the composition according to the present invention, for example, a pharmaceutical composition, is formulated by using a pharmaceutically acceptable carrier according to a method that can be clearly practiced by those skilled in the art to which the present invention pertains. It can be prepared in dosage form or placed into a multi-dose container. In the present invention, the term 'carrier' means a compound that facilitates the addition of a compound into cells or tissues, and the term 'pharmaceutically acceptable' means that it is physiologically acceptable and when administered to humans, it is usually gastrointestinal. It refers to a composition that does not cause allergic reactions such as disability or dizziness or similar reactions.

The pharmaceutically acceptable carrier is one commonly used in formulation, and includes lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum acacia, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, and polyvinyl. pyrrolidone, cellulose, water, syrup, methylcellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil, but are not limited thereto.

In addition, the composition according to the present invention, for example, a pharmaceutical composition, may further include additives such as a filler, an anti-agglomerating agent, a lubricant, a wetting agent, a flavoring agent, an emulsifier, and a preservative, in addition to the above components. In the present invention, the content of the additives included in the composition is not particularly limited and may be appropriately adjusted within the range of content used in conventional formulations.

In addition, the composition according to the present invention, for example, a pharmaceutical composition, a food composition, or a probiotics composition may be formulated as an oral preparation. Non-limiting examples of the oral preparations include tablets, troches, lozenges, aqueous suspensions, oily suspensions, prepared powders, granules, emulsions, hard capsules, soft capsules, syrups or elixirs. can be heard In order to formulate the pharmaceutical composition according to the present invention for oral administration, a binder such as lactose, saccharose, sorbitol, mannitol, starch, amylopectin, cellulose or gelatin; excipients such as dicalcium phosphate and the like; disintegrants such as corn starch or sweet potato starch; Magnesium stearate, calcium stearate, sodium stearyl fumarate, and the like can be used, and sweeteners, aromatics, syrups, and the like can also be used. Furthermore, in the case of capsules, a liquid carrier such as fatty oil may be additionally used in addition to the above-mentioned materials.

In the present invention, the term 'excipient' refers to any substance other than a therapeutic agent, and is used as a carrier or medium for delivery of a therapeutic agent or added to a pharmaceutical composition. This improves handling and storage properties or allows and facilitates unit dosage formation of the composition.

The composition according to the present invention, for example, a pharmaceutical composition, a food composition, or a probiotics composition, according to a conventional method according to each purpose of use, is a liquid, suspension, powder, granules, tablets, capsules, pills, extracts, emulsions It can be formulated and used in various forms, such as oral formulations such as syrups and aerosols, and injections of sterile injection solutions, and can be administered orally or through various routes including intravenous, intraperitoneal, subcutaneous, rectal, and topical administration. there is. In the present invention, the term 'oral administration' means a substance prepared to digest an active substance, that is, administered to the gastrointestinal tract for absorption.

A preferred dosage of the composition according to the present invention, for example, a pharmaceutical composition, food composition, or probiotics composition, depends on the condition and weight of the patient, age, sex, health condition, dietary constitutional specificity, the nature of the preparation, the severity of the disease, and the composition of the composition. The range may vary depending on the administration time, administration method, administration period or interval, excretion rate and drug type, and may be appropriately selected by a person skilled in the art.

In the present invention, the term 'effective dose of the pharmaceutical composition' means an amount of the composition of the active ingredient sufficient to treat a specific symptom. This may vary depending on the formulation method, administration method, administration time and/or route of administration of the pharmaceutical composition, and the type and degree of response to be achieved by administration of the pharmaceutical composition, the type of subject to be administered, age, Weight, general state of health, symptoms or severity of disease, sex, diet, excretion, drugs or other components of the composition used simultaneously or concurrently in the individual, and similar factors well known in the medical arts. And those skilled in the art can easily determine and prescribe an effective dosage for the desired treatment.

Administration of the pharmaceutical composition according to the present invention may be administered once a day, or may be divided and administered several times. The composition may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents. Considering all of the above factors, it can be administered in an amount that can obtain the maximum effect with the minimum amount without side effects.

For example, the composition according to the present invention is 0.001 to 10,000 mg, 0.001 to 5,000 mg, 0.001 to 1,000 mg, 0.001 to 500 mg, 0.001 to 300 mg, 0.001 to 100 mg, 0.001 to 50 mg, 0.001 to 1 kg of body weight. to 30 mg, 0.001 to 10 mg, 0.001 to 5 mg, 0.001 to 1 mg, 0.001 to 0.5 mg, 0.001 to 0.1 mg, 0.001 to 0.05 mg, 0.001 to 0.01 mg, 0.01 to 10,000 mg, 0.01 to 5,000 mg , 0.01 to 1,000 mg, 0.01 to 500 mg, 0.01 to 300 mg, 0.01 to 100 mg, 0.01 to 50 mg, 0.01 to 30 mg, 0.01 to 10 mg, 0.01 to 5 mg, 0.01 to 1 mg, 0.01 to 0.5 mg, 0.01 to 0.1 mg, 0.01 to 0.05 mg, 0.1 to 10,000 mg, 0.1 to 5,000 mg, 0.1 to 1,000 mg, 0.1 to 500 mg, 0.1 to 300 mg, 0.1 to 200 mg, 0.1 to 100 mg, 0.1 to 50 mg, 0.1 to 30 mg, 0.1 to 10 mg, 0.1 to 5 mg, 0.1 to 1 mg, 0.1 to 0.5 mg, 1 to 10,000 mg, 1 to 5,000 mg, 1 to 1,000 mg, 1 to 500 mg, 1 to 300 mg, 1 to 200 mg, 1 to 100 mg, 1 to 50 mg, 1 to 10 mg, 1 to 5 mg, 10 to 10,000 mg, 10 to 5,000 mg, 10 to 1,000 mg, 10 to 500 mg, 10 to 300 mg, 10 to 200 mg, 10 to 100 mg, 10 to 50 mg, 10 to 40 mg, 10 to 30 mg, 10 to 20 mg, 100 to 10,000 mg, 100 to 5,000 mg, 100 to 1,000 mg, 100 to 500 mg, 100 to 300 mg, or 100 to 200 mg per day, but is not limited thereto. For example, the daily dose of the composition according to the present invention is 0.001 to 10 g/day, 0.001 to 5 g/day, 0.01 to 10 g/day, or 0.01 to 5 g/day based on oral administration to adult patients. It may be 1 day. In addition, the total daily dose can be divided and administered continuously or discontinuously as needed.

In the present invention, the term “culture of a strain” refers to a product obtained after culturing a strain according to an example of the present invention, and the culture includes the entire culture of the strain according to an example of the present invention, a dilution thereof, It may be a concentrate, dried product, freeze-dried product, lysate, and/or fraction, etc., and the concentrate may be obtained by centrifuging or evaporating the culture, and the dried product may be obtained by drying the culture using a dryer or the like. It can be obtained by drying, and the freeze-dried product can be obtained by freeze-drying the culture using a freeze dryer, etc., and the lysate can be obtained by physically or sonicating the strain or culture. Fractions can be obtained by applying methods such as centrifugation and chromatography to the cultures and lysates. The culture may be in a solid phase (solid, eg, dry matter), liquid phase (liquid), or fluidized phase, but is not limited thereto. In one example, the culture may mean a cultured strain obtained by culturing a strain according to an embodiment of the present invention for a certain period of time, a metabolite thereof, and/or a whole medium including extra nutrients. In one example, the culture may be one in which the strain according to an example of the present invention has not been removed or removed. In one example, the culture may mean the remaining components except for the strain (cell) in the culture in which the strain according to an embodiment of the present invention is cultured in a medium. In one example, the culture may be a culture medium (or culture) in which the strain (cell) is removed from the culture medium in which the strain according to an embodiment of the present invention is cultured in a medium. The culture solution (or culture product) from which the strain is removed may be a cell free culture solution (or culture product) or a culture solution containing dead cells, and for example, a filtrate from which the strain is removed by filtration or centrifugation supernatant), and/or a culture medium containing dead cells (or a dried material of the culture medium). Specifically, the culture may exhibit an anti-inflammatory activity equivalent to that of the strain according to an embodiment of the present invention, or an activity for preventing, improving or treating inflammatory diseases. For example, the culture may contain extracellular polysaccharide produced by the strain according to an embodiment of the present invention.

In the present invention, the term "lysate of a strain" may mean a product obtained by crushing a strain according to an embodiment of the present invention by chemical or physical force. Specifically, the lysate may exhibit an anti-inflammatory activity equivalent to the activity exhibited by the strain according to an embodiment of the present invention, or an activity for preventing, improving or treating inflammatory diseases. For example, the lysate may include extracellular polysaccharide produced by the strain according to an embodiment of the present invention.

In the present invention, the term "extract" refers to extracting a strain according to an example of the present invention, a culture of the strain, a lysate of the strain, or a mixture thereof regardless of the extraction method, extraction solvent, extracted component, or extract form. It can mean the obtained product, and is a broad concept that includes all substances that can be obtained by processing or processing by other methods after extraction. For example, the extract may be an extract of a strain according to an embodiment of the present invention, an extract of a culture of the strain, or an extract of a lysate of the strain. Specifically, the extract may exhibit anti-inflammatory activity equivalent to that of the strain, the culture of the strain, or the lysate of the strain according to an embodiment of the present invention, or an activity for preventing, improving or treating inflammatory diseases. there is. For example, the extract may contain extracellular polysaccharide produced by the strain according to an embodiment of the present invention.

The Picalibacterium prausnitzii strain according to an example of the present invention is superior in promoting anti-inflammatory cytokine production compared to conventional strains, exhibits anti-inflammatory properties, and exerts a therapeutic effect on inflammatory diseases, thereby preventing, improving, and treating inflammatory diseases It can be used in compositions for

Figure 1a is a diagram showing the results of intestinal flora analysis of KO-16 subjects from which P. prausnitzii KBL1027 strain was isolated and KO-13 subjects from whom KBL1026 strain was isolated based on 16S rRNA.
Figure 1b is a diagram showing the results of analyzing the distribution of intestinal flora of KO-16 subjects from which P. prausnitzi KBL1027 strain was isolated and KO-13 subjects from which KBL1026 strain was isolated.
Figure 2a is a diagram showing the change in body weight according to the administration of the Picalibacterium prausnitzii strain according to an example of the present invention.
Figure 2b is a diagram showing the change in the length of the large intestine according to the administration of the Picalibacterium prausnitzii strain according to an example of the present invention.
Figure 2c is a view showing the process of measuring the change in the length of the large intestine according to the administration of the Picalibacterium prausnitzii strain according to an example of the present invention.
3 is a view showing the results of H&E staining after administration of the Picalibacterium prausnitzii strain according to an example of the present invention.
Figure 4a is a diagram showing changes in the expression level of the anti-inflammatory cytokine gene IL-10 according to the administration of the Picalibacterium prausnitzii strain according to an example of the present invention.
Figure 4b is a diagram showing changes in the expression level of the tight junction gene Zo-1 according to the administration of the Picalibacterium prausnitzii strain according to an example of the present invention.
Figure 4c is a diagram showing the change in the expression level of the tight junction gene Occludin according to the administration of the Picalibacterium prausnitzii strain according to an example of the present invention.
Figure 5a is a diagram showing changes in the diversity of intestinal flora according to the administration of the Picalibacterium prausnitzii strain according to an example of the present invention.
5B is a view showing the main component analysis results through the weighted UniFrac distance of the intestinal flora after administration of the Picalibacterium prausnitzii strain according to an example of the present invention.
Figure 5c is a diagram showing the results of univariate analysis (LefSE) to determine the change in intestinal flora according to the administration of the Picalibacterium prausnitzii strain according to an example of the present invention.
Figure 6 is a view showing the results of observing the Picalibacterium prausnitzii strain with a scanning electron microscope.
7 is a diagram showing the anti-inflammatory effect of the extracellular polysaccharide specifically produced by the Picalibacterium prausnitzi KBL1027 strain.
8 is a diagram showing genes specifically present in the genome of strain KBL1027 of Picalibacterium prausnitzi according to their functional characteristics.
9 is a view showing genes belonging to the cell wall and capsule function specifically present in the genome of the P. prausnitzii KBL1027 strain.

Hereinafter, the present invention will be described in more detail by the following examples. However, these examples are only for illustrating the present invention, and the scope of the present invention is not limited by these examples.

Example 1. Isolation of new strains

Novel Faecalibacterium prausnitzii strains were isolated from the intestinal flora of Koreans. Specifically, samples for intestinal flora isolation were isolated and identified from fecal samples provided from different healthy adults who had not taken antibiotics for 6 months (Seoul National University IRB approval number: 1602/001-001). Immediately after collection, fecal samples were transported to our laboratory and immediately transferred to an anaerobic chamber (Coy Laboratory Products Inc.) to be used for strain isolation. The samples were streaked in YCFAG medium containing 1.5% agar in a direct smear format, and then incubated for up to 48 hours at 37 °C under anaerobic conditions. After culturing, pure isolated colonies were randomly selected and cultured in YBHI medium. For long-term storage of strains, glycerol (25% v/v) was added to the culture medium that reached the exponential phase and stored in -80 ° C cryogenic freezer. kept. To identify the strain, extract the genomic DNA of the strain using the Wizard genomic purification kit (Promega), and then perform a PCR reaction using the 27F/1492R primer (SEQ ID NO: 1 and 2) in Table 1 below targeting the 16S rRNA gene did

division Sequence (5'→ 3') sequence number forward AGAGTTTGATYMTGGCTCAG One reverse TACGGYTACCTTGTTACGACT 2

After purification using the QIAquick PCR purification kit (Qiagen), sequencing was performed using the ABI3711 automatic sequencer (Macrogen). The 16S rRNA analysis results are shown in Table 2 below, and using this sequence information, multiple comparisons were made with Cheonlab's EzBioCloud program (http://www.ezbiocloud.net/identify) to finally complete the strain identification.

strain name denomination order sequence number Faecalibacterium prausnitzii KBL1026 GACGAACGCTGGCGGCGCGCCTAACACATGCAAGTCGAACGAGTGAGAGAGCTTGCTTTCTCGAGCGAGTGGCGAACGGGTGAGTAACGCGTGAGGAACCTGCCTCAAAGAGGGGGACAACAGTTGGAAACGACTGCTAATACCGCATAAGCCCACGACCCGGCATCGGGTAGAGGGAAAAGGAGCAATCCGCTTTGAGATGGCCTCGCGTCCGATTAGCTAGTTGGTGAGGTAACT GGCCCACCAAGGCGACGATCGGTAGCCGGACTGAGAGGTTGAACGGCCACATTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGGGAAACCCTGATGCAGCGACGCCGCGTGGAGGAAGAAGGTCTTCGGATTGTAAACTCCTGTTGTTGAGGAAGATAATGACGGTACTCAACAAGGAAGTGACGGCTAACTACGTGCCAGCAGCCGC GGTAAAACGTAGGTCACAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGCAGGCGGGAAGACAAGTTGGAAGTGAAATCCATGGGCTCAACCCATGAACTGCTTTCAAAACTGTTTTTCTTGAGTAGTGCAGAGGTAGGCGGAATTCCCGGTGTAGCGGTGGAATGCGTAGATATCGGGAGGAACACCAGTGGCGAAGGCGGCCTACTGGGCACCAACTGACGCTGAGGCTCGAAAGTGTG GGTAGCAAACAGGATTAGATACCCTGGTAGTCCACACTGTAAACGATGATTACTAGGTGTTGGAGGATTGACCCCTTCAGTGCCGCAGTTAACAATAAGTAATCCACCTGGGGAGTACGACCGCAAGGTTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCAGTGGAGTATGTGGTTTAATTCGACGCAACGCGAAGAACCTTACCAAGTCTTGACATCCTGCGACGCACATAGAAACA GTAGTTTCCTTCGGGACGCAGAGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTATGGTCAGTTACTACGCAAGAGGACTCTGGCCAGACTGCCGTTGACAAAACGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCTTTATGACTTGGGCTACACACGTACTACAATGGCGTTAAACAAAGAGAAGCAAGACCGA GGTGGAGCAAAACTCAGAAACAACGTCCCAGTTCGGACTGCAGGCTGCAACTCGCCTGCACGAAGTCGGAATTGCTAGTAATCGCAGATCAGCATGCTGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGAGAGCCGGGGGGACCCGAAGTCGGT 3 Faecalibacterium prausnitzii KBL1027 TGAATTTGGCTCAGGACGAACGCTGGCGGCGCGCCTAACACATGCAAGTCGAACGAGCGAGAGAGCTTGCTTTCTCAGAGCGAGTGGCGAACGGGTGAGTAACGCGTGAGGAACCTGCCTCAAAGAGGGGGACAACAGTTGGAAACGACTGCTAATACCGCATAAGCCCACGACCCGGCATCGGGTAGAGGGAAAAGGAGCAATCCGCTTTGAGATGGCCTCGCGTCCGATTAGCT AGTTGGTGAGGTAACGGCCCACCAAGGCGACGATCGGTAGCCGGACTGAGAGGTTGAACGGCCACATTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGGGAAACCCTGATGCAGCGACGCCGCGTGGAGGAAGAAGGTCTTCGGATTGTAAACTCCTGTTGTTGAGGAAGATAATGACGGTACTCAACAAGGAAGTGACGGCTAACT ACGTGCCAGCAGCCGCGGTAAAACGTAGGTCACAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGCAGGCGGGAAGACAAGTTGGAAGTGAAATCCATGGGCTCAACCCATGAACTGCTTTCAAAACTGTTTTTCTTGAGTAGTGCAGAGGTAGGCGGAATTCCCGGTGTAGCGGTGGAATGCGTAGATATCGGGAGGAACACCAGTGGCGAAGGCGGCCTACTGGGCACCAACTGACGCTG AGGCTCGAAAGTGTGGGTAGCAAACAGGATTAGATACCCTGGTAGTCCACACTGTAAACGATGATTACTAGGGGTTGGAGGATTGACCCCTTCAGTGCCGCAGTTAACACAATAAGTAATCCACCTGGGGAGTACGACCGCAAGGTTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCAGTGGAGTATGTGGTTTAATTCGACGCAACGCGAAGAACCTTACCAAGTCTTGACATCCTGC GACGCACATAGAAATATGTGTTTCCTTCGGGACGCAGAGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTATGGTCAGTTACTACGCAAGAGGACTCTGGCCAGACTGCCGTTGACAAAACGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCTTTATGACTTGGGCTACACGTACTACAATGGCGTTAAACAAAGA GAAGCAAGACCGCGAGGTGGAGCAAAACTCAGAAACAACGTCCCAGTTCGGACTGCAGGCTGCAACTCGCCTGCACGAAGTCGGAATTGCTAGTAATCGCAGATCAGCATGCTGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGAGAGCCGGGGGGACCCGAAGTCGGTAGTCTAACCGCAAGGAG 4

The two isolated new strains were named as Picalibacterium prausnitzii KBL1026 strain and Picalibacterium prausnitzii KBL1027 strain, respectively, and deposited at the Korea Research Institute of Bioscience and Biotechnology Biological Resource Center on July 7, 2020, and each deposit number KCTC14230BP (Picalibacterium prausnitzii KBL1026) and KCTC14231BP (Picalibacterium prausnitzii KBL1027) were assigned.

Example 2. Structural analysis of the intestinal microbiota of subjects with new strains

The new strain, Picalibacterium prausnitzii KBL1026 strain, which is a new strain isolated in Example 1, was derived from a KO-13 subject, and the Phicalibacterium prausnitzii KBL1027 strain was derived from a KO-16 subject. In order to analyze the intestinal flora structure of subjects with new strains, the obtained stool samples were stored in a cryogenic freezer at -80 ° C for the purpose of intestinal flora analysis, and the frozen samples were transferred to the laboratory to use QIAamp FAST DNA stool mini kit (Qiagen) was used to extract total bacterial genomic DNA in feces. The extracted DNA was amplified using 515F/806R primers (SEQ ID NOs: 5 and 6) in Table 3 below targeting the V4 region of the bacterial 16S rRNA gene, and then sequencing data was generated using MiSeq (Illumina). . The generated large-capacity sequencing analysis identified the structure of the intestinal flora by confirming the entire genetic information of the intestinal bacteria using the QIIME pipeline.

division Sequence (5'→ 3') sequence number forward ATGATACGGCGACCACCGAGATCTACACTATGGTAATTGTGTGCCAGCMGCCGCGGTAA 5 reverse CAAGCAGAAGACGGCATACGAGATAGTCAGTCAGCCGGACTACHVGGGTWTCTAAT 6

As a result of analyzing the intestinal flora based on 16S rRNA, as shown in FIG. 1a, the KO-16 subjects from which the P. prausnitzi KBL1027 strain was isolated had a higher intestinal flora than the KO-13 subjects from which the KBL1026 strain was isolated. It was confirmed that the diversity of was high (Fig. 1a).

In addition, as a result of analyzing the distribution of bacteria occupying a ratio of 1% or more at the species level of the flora, as shown in FIG. It was confirmed that the relative abundance) was dominant as high as 15.5% or more (Fig. 1b). On the other hand, it was confirmed that the KO-13 subjects from whom the KBL1026 strain was isolated occupied a very low ratio of 0.8% in the relative abundance of the intestinal P. prausnitzii strain (FIG. 1b).

The difference in the relative abundance of P. prausnitzi in the intestines of these subjects is due to the fact that the newly acquired P. prausnitzi KBL1026 and KBL1027 strains belong to the same species, but the intestinal flora environment of the derived subjects is different. It was suggested that the functionality may be different.

Example 3. Inflammatory disease improvement effect analysis using animal models

(1) Animal model establishment and diet

Animal experiments were conducted to confirm the effect of improving inflammatory diseases in vivo by administering a single strain of P. prausnitzii KBL1026 or KBL1027 strain (Seoul National University IACUC approval number: SNU-160602-9). As an example of an inflammatory disease, an inflammatory bowel disease animal model was used for experiments. In order to compare the inflammatory disease improvement efficacy of the strains, the Phycalibacterium prausnitzii DSM17677 strain belonging to the same species and known to have anti-inflammatory efficacy was distributed from the German Strains Bank (DSMZ) and used as a control group.

Specifically, in order to construct a colitis mouse model, C57BL/6 7-8 week-old female mice were divided into groups of 8 each, and 2.5% dextran sulfate sodium (DSS) was dissolved in drinking water for a total of 5 days. Acute enteritis was induced by drinking. The normal control group was supplied with drinking water to which DSS was not added.

After culturing the P. prausnitzi KBL1026, KBL1027, or DSM17677 strains in YBHI liquid medium at 37 ° C. in anaerobic conditions to reach the exponential phase, remove the supernatant and dilute in PBS to 2x10 ⁸ CFU / ml, From one week before the start of DSS supply until the end of the experiment, 200 μl of each of the diluted strains was orally administered to mice every day for colonization. A normal control group was orally administered 200 μl of PBS daily.

(2) Weight change analysis

DSS supply was stopped after 5 days (Day 5 morning), and the weight change of the mice was measured every day for 9 days from the start day of DSS supply (Day 0), and is shown in FIG. 2a. Figure 2a is a view showing changes in body weight of mice according to acute enteritis. As shown in Figure 2a, compared to the normal control group (Water + PBS) not administered with DSS, the DSS control group (DSS + PBS) showed a significant weight loss, confirming the construction of the colitis mouse model. In addition, the KBL1027 administration group (DSS + KBL1027) confirmed a significantly improved effect on weight loss compared to the DSS control group (DSS + PBS). On the other hand, the KBL1026-administered group and the DSM17677-administered group had no significant weight change compared to the DSS control group (DSS+PBS).

(3) Chapter length analysis

On the ninth day after the start of DSS supply, the experiment was terminated, the mice were necropsied, and the change in intestinal length was measured and shown in FIG. 2B. As shown in Figure 2b, it was confirmed that the KBL1027-administered group significantly improved the reduction in colon length compared to the DSS control group. Figure 2c is a view showing the process of measuring the intestinal length after autopsy of the mouse.

(4) Histopathological analysis of the intestine

In addition, Hematoxylin & Eosin (H&E) staining was performed to observe histopathological changes in the colon due to the administration of a single strain of P. prausnitzii. Specifically, after autopsy, the distal part of the large intestine was fixed in 10% neutral formalin solution, and paraffin tissue specimens were sectioned at a thickness of 5 μm, stained with H&E reagent, and observed under an optical microscope. The H&E staining results are shown in FIG. 3, and in the DSS control group (DSS + PBS), compared to the normal control group (Water + PBS), infiltration of inflammatory cells in the colon tissue and significant destruction of the mucosal tissue were observed. On the other hand, it was confirmed histologically that the KBL1027 administration group (DSS+KBL1027) had reduced inflammation compared to the DSS control group (DSS+PBS).

(5) Analysis of inflammatory disease improvement effect in vivo using marker genes

For tissue immune cell analysis, mouse intestinal tissue was obtained at the end of the experiment in Example 2, preserved in RNAlater (Thermo Fisher Scientific) solution and stored frozen at -81 ° C, and easy-spin total RNA extraction kit to extract RNA (Intron) was used. The extracted RNA was immediately synthesized into cDNA using the high capacity RNA-to-cDNA kit (Applied biosystems) in the same amount, and then the gene expression level was analyzed using the roter-gene SYBR green PCR kit (Qiagen). The primers in Table 4 were used to target IL-10, an anti-inflammatory cytokine, Zo-1 related to intestinal epithelial tight junction, and Occludin genes, and the expression level as an HPRT house keeping gene. 4a (IL-10), 4b (Zo-1), and 4c (Occludin) show the gene expression level analysis results by correcting .

division target gene Sequence (5'→ 3') sequence number forward HPRT TTATGGACAGGACTGAAAGAC 7 reverse HPRT GCTTTAATGTAATCCAGCAGGT 8 forward IL-10 ATAACTGCACCCACTTCCCA 9 reverse IL-10 TCATTTCCGATAAGGCTTGG 10 forward Zo-1 ACCCGAAACTGATGCTGTGGATAG 11 reverse Zo-1 AAATGGCCGGGCAGAACTTGTGTA 12 forward Occludin GGAGGACTGGGTCAGGGAATA 13 reverse Occludin CGTCGTCTAGTTCTGCCTGT 14

As shown in Figure 4a, the expression level of the anti-inflammatory cytokine IL-10 gene was significantly increased in the KBL1027 administration group (DSS + KBL1027) compared to the DSS control group (DSS + PBS). This means that the Picalibacterium prausnitzi KBL1027 strain contributes to the improvement of inflammatory diseases in vivo through immune regulation that induces IL-10 production.

According to the results of FIGS. 4b and 4c, in the case of Zo-1 and Occludin related to the tight junction between intestinal epithelial cells, both genes significantly decreased in the DSS control group (DSS + PBS) compared to the normal control group. seemed On the other hand, compared to the DSS control group (DSS + PBS), the KBL1027 administration group (DSS + KBL1027) significantly increased the expression levels of both Zo-1 and Occludin . This means that administration of the KBL1027 strain increased the expression levels of Zo-1 and Occludin , strengthened the tight junctions between intestinal epithelial cells, and helped alleviate inflammatory diseases in vivo.

Example 4. Analysis of changes in intestinal flora using animal models

In order to analyze changes in intestinal flora induced by colonization of a single strain and acute enteritis caused by DSS, feces of mice were obtained at the end of the experiment in Example 3 and stored frozen at -81 ° C. Total bacterial genomic DNA was extracted from the frozen samples according to the method of Example 1, and large-capacity sequencing data were generated targeting the V4 region of the bacterial 16S rRNA gene. The total genetic information of intestinal bacteria was confirmed using the QIIME pipeline to determine the structure of the flora in mouse feces, and univariate analysis (LefSE) was performed according to the group, and the results are shown in FIGS. 5A to 5C.

The results of confirming the diversity change of the intestinal flora analyzed based on 16S rRNA by Faith's Phylogenetic diversity index according to the group are shown in Figure 5a. ), it was confirmed that the diversity of intestinal flora was significantly reduced. This suggests that acute enteritis caused by DSS can negatively affect intestinal health due to the decrease in the diversity of beneficial bacteria and the dominance of potential harmful bacteria. On the other hand, the diversity of intestinal flora increased significantly in the KBL1027-administered group (DSS + KBL1027) compared to the DSS control group (DSS + PBS), and there was no significance in the DSM17677-administered group (DSS + DSM17677). This means that even the same strain of Ficcallybacterium prausnitzii has different effects on the intestinal flora, and administration of the Ficcallybacterium prausnitzii KBL1027 strain restores the diversity of the intestinal flora and has a positive effect on intestinal health. suggests that it can

The results of principal component analysis through weighted UniFrac distance of the intestinal flora analyzed based on 16S rRNA are shown in Figure 5b, and the PCA plot confirmed that the DSS control group had a very different intestinal flora structure from the normal control group. On the other hand, it was confirmed that the KBL1027-administered group had an intestinal flora structure between the DSS control group and the normal control group. This suggests that administration of the KBL1027 strain modulated the structure by changing the species constituting the intestinal flora.

Figure 5c shows the results of univariate analysis (LefSE) to analyze which intestinal flora was changed by the administration of KBL1027 strain. In the DSS control group, the dominance of Bacteroides and Proteobacteria, whose cell wall is composed of lipopolysaccharide (LPS), which is a representative harmful bacterium, was confirmed. Meanwhile, in the KBL1027-administered group, an increase in Prevotella and Paraprevotellaceae was confirmed. This means that administration of the KBL1027 strain helped alleviate colitis by changing the intestinal flora.

Example 5. F. prausnitzii Anti-inflammatory test of substances derived from KBL1027 strain

(1) Observation of extracellular polysaccharide specific to strain KBL1027

In order to confirm the strain specificity between the P. prausnitzi strains, the three-dimensional surface morphology of the bacterial samples was observed using a scanning electron microscope (SEM).

Specifically, samples obtained by incubating P. prausnitzii KBL1026, KBL1027, and DSM17677 strains in YBHI medium at 37 ° C. under anaerobic conditions for 48 hours were fixed using karnovsky's solution and 2% osmium tetroxide, ethanol dehydrated, and critical point A pretreatment process of drying was performed. Thereafter, it was fixed to a sample stand (stub), deposited with platinum (coating), and observed using a scanning electron microscope (Carl Zeiss). Observation photographs are shown in FIG. 6 , and in the case of KBL1027, it was confirmed that extracellular polysaccharide material was produced on the cell surface in a strain-specific manner compared to the KBL1026 and DSM17677 strains. On the other hand, in the case of KBL1026, a form similar to that of DSM17677 reported previously was observed. As such, the material specifically produced by the KBL1027 strain was confirmed through the observation of the bacterial cell surface, and this strain specificity is the outstanding characteristic of the KBL1027 strain compared to other strains belonging to the Ph. It suggests that inflammatory disease alleviation function can be derived.

(2) Anti-inflammatory effect of KBL1027 strain by extracellular polysaccharide

To verify the immunomodulatory effect of substances produced by the P. prausnitzii KBL1027 strain, mouse bone-marrow derived macrophage (BMDM) was isolated and P. prausnitzi KBL1026, KBL1027 , And by processing the culture medium of the DSM17677 strain to confirm the ability to produce IL-10, a cytokine involved in the anti-inflammatory response.

Specifically, in order to obtain a culture solution of the strain, Ph. calibacterium prausnitzi KBL1026, KBL1027, and DSM17677 strains were cultured in 500 ml of YBHI medium at 37 ° C. under anaerobic conditions for 48 hours to obtain a supernatant through centrifugation and a 0.45 μm filter proceed to. As a negative control, a YBHI medium not inoculated with the strain was used, and the obtained medium and supernatant were frozen at -81 ° C, then lyophilized and diluted in PBS in the same amount to 100 mg / ml.

To obtain mouse BMDM cells, the bone marrow of C57BL/6 mice was separated, and 10% fetal bovine serum (FBS), 15% L292 cell culture medium, and 1% penicillin/streptomycin antibiotics were added to DMEM medium. It was cultured for 7 days in a medium for BMDM using an incubator at 37 °C and 5% CO ₂ conditions. L292 cells were purchased from the Korean Cell Line Bank.

BMDM cells of the mouse were dispensed into each well of a 96-well plate by 2×10 ⁴ cells and attached for 24 hours, and then the negative control YBHI medium and the supernatant of the P. prausnitzi KBL1026, KBL1027, and DSM17677 strains were added to the 1 It was replaced by adding it to the culture medium at a concentration of mg/ml. In addition, in order to confirm the reaction upon stimulation of the inflammatory response induced by LPS, LPS was added at a concentration of 100 ng/ml, and then the supernatant of each strain was added in the same amount. As a positive control, LPS was treated at concentrations of 10 ng/ml and 100 ng/ml, and sodium butyrate (sigma aldrich) was treated at 1 ng for comparison with the butyric acid metabolite produced by Picalibacterium prausnitzii. /ml, 10 ng/ml. After 24 hours had elapsed, the culture medium was collected and stored frozen at -81 ° C. Then, the amount of IL-10 cytokine was measured using a mouse IL-10 ELISA kit (Thermo Fisher Scientific) according to the manufacturer's method.

The results are shown in FIG. 7, and in the BMDM cells of the mouse, the culture medium of the P. prausnitzii KBL1027 strain was most significantly anti-inflammatory cytokine IL compared to the negative control YBHI medium and the culture medium of the KBL1026 and DSM17677 strains. -10 was found to induce production. In addition, even when the inflammatory response was stimulated by LPS, IL-10 production was induced more significantly than the negative control YBHI medium and the culture medium of KBL1026 strain. These experimental results indicate that the substances specifically produced and excreted by the P. prausnitzii KBL1027 strain induce the production of the anti-inflammatory cytokine IL-10.

Example 6. F. prausnitzii Comparative genomic analysis of strain KBL1027

Comparative genomic analysis was performed to confirm the strain specificity between the strains of Picalibacterium prausnitzii. In order to secure the bacterial genome information, KBL1026 and KBL1027 strains were cultured in YBHI medium at 37 ° C. under anaerobic conditions, and genomic DNA of the strains was extracted according to the method of Example 1. In the case of KBL1026, large-capacity sequencing data were generated using MiSeq (Illumina) and assembled according to the A5-MiSeq pipeline, and the results are shown in Table 5 below. In the case of KBL1027, complete genome information was obtained using RSII (PacBio). Genomic information of the DSM17677 strain was obtained from NCBI and used for comparative genomic analysis (accession number: GCA_000162015.1). For comparison of the average nucleotide identity (ANI) of the genome, it was calculated through JSpeciesWS (http://jspecies.ribohost.com/jspeciesws/#analyse), and the summary and comparison of the genome information of the obtained strains are shown. shown in 5. The ANI values of strains KBL1027 and KBL1026 were 96.13%, whereas those of strain DSM17677 were 83.25%. This means that the KBL1027 strain has a higher genome sequence similarity with the KBL1026 strain than the previously known DSM17677 strain.

division KBL1026 KBL1027 DSM17677 Size (bp) 3,105,066 3,124,218 3,090,349 GC content (%) 56.2 56.1 56.4 Contig N50 (bp) 330,297 - 63,061 Number of scaffolds 36 One 20 Number of coding sequences 3,193 3176 3,298 Number of RNAs 71 83 79 ANI (%) with KBL1027 96.13 100 83.25

Based on the obtained genome information of the Picalibacterium prausnitzii strain, RAST (https://rast.nmpdr.org/) was used to perform annotation along the RASTtk pipeline and comparative genomic analysis was performed. A total of 61 genes were specifically present in the genome of strain KBL1027 compared to strain DSM17677, and a total of 15 genes were specifically present in genome of strain KBL1027 compared to strain KBL1026. This is interpreted as the fact that more genes specifically exist because the KBL1027 strain has a lower ANI value than the DSM17677 strain. When compared with the two comparative strains, KBL1026 strain and DSM17677 strain, respectively, the number of genes specifically present only in the genome of KBL1027 strain is shown in FIG. 8 according to their functional characteristics.

Among them, in the case of cell wall and capsule functions related to the production of extracellular polysaccharide substances revealed in Example 5, 7 genes compared to KBL1026 strain and 10 genes compared to DSM17677 strain were found in KBL1027 strain. It was confirmed that it exists specifically.

Next, 13 genes (below (1) to (13)), which were absent in both KBL1026 and DSM17677 strains and specifically existed only in the KBL1027 genome, were identified, of which 7 (below (1) to ( 7)) was confirmed to belong to the Cell Wall and Capsule function:

(1) Exopolysaccharide biosynthesis glycosyltransferase EpsF (EC 2.4.1.-)

(2) Tyrosine-protein kinase EpsD (EC 2.7.10.2)

(3) Alpha-D-GlcNAc alpha-1,2-L-rhamnosyltransferase (EC 2.4.1.-) rgpA

(4) Alpha-L-Rha alpha-1,3-L-rhamnosyltransferase (EC 2.4.1.-) rgpB

(5) capsular polysaccharide biosynthesis protein

(6) Lipoprotein releasing system ATP-binding protein LolD

(7) Putative N-acetylgalactosaminyl-diphosphoundecaprenol glucuronosyltransferase TuaG

(8) Predicted transcriptional regulator of N-Acetylglucosamine utilization, GntR family NagQ

(9) Membrane-bound lytic murein transglycosylase B (EC 3.2.1.-)

(10) Inosine-uridine preferring nucleoside hydrolase (EC 3.2.2.1)

(11) LysR family transcriptional regulator YnfL

(12) Cation efflux system protein CusA

(13) Cobalt-zinc-cadmium resistance protein CzcA

The genes belonging to the cell wall and capsule functions specifically present in the genome of the P. prausnitzi KBL1027 strain are shown in FIG. 9, and the KBL1027 strain-specific genes are exopolysaccharide biosynthesis (exopolysaccharide biosynthesis) and rhamnose containing glycans, and it was confirmed that they form a cluster (FIG. 9B). Figure 9B is a diagram showing the results of confirming the cluster of genes 1, 3, 4, and 5 in A of Figure 9 on the genome sequence of strain KBL1027 of Picalibacterium prausnitzii. Such comparative genomic analysis results indicate that the P. prausnitzii KBL1027 strain specifically produces an extracellular polysaccharide material.

Example 7. F. prausnitzii Evaluation of in vitro anti-inflammatory activity of strain KBL1027

(1) NO assay

After inducing inflammation by treating mouse macrophage RAW264.7 cells with LPS, the amount of NO production was measured by treating the P. prausnitzii KBL1027 strain according to an example of the present invention, thereby confirming anti-inflammatory activity.

(2) Cytokine measurement

After inducing inflammation by treating mouse macrophage RAW264.7 cells with LPS, the cytokine TNF-a, IL-10 and Anti-inflammatory activity was confirmed by measuring IL-6. Among cytokines, it was confirmed that the expression of anti-inflammatory cytokine IL-10 increased, and the inflammatory mediating cytokines TNF-a and IL-6 decreased compared to the control group.

TNF-a is an inflammation-inducing cytokine that induces the migration of neutrophil monocytes and lymphocytes in blood vessels, upregulates the production of collagenase and PGE2, and is involved in osteoclast differentiation.

IL-10 is a cytokine secreted by Th2 cells that suppresses inflammation and allergy.

IL-6 is involved in inflammatory responses, host responses and tissue damage, and is secreted in large amounts by inflammatory stimuli such as IL-1 and TNFa. It promotes osteoclast differentiation and causes bone loss.

In addition, anti-inflammatory activity was confirmed by measuring COX-2 and iNOS through immunoblot analysis.

COX-2 is a protein induced by LPS that promotes an acute inflammatory response.

INOS produces NO by external stimuli or inflammatory cytokines.

Example 8. F. prausnitzii Evaluation of anti-allergic activity of strain KBL1027

(One) β-hexosaminidase assay (measuring absorbance)

β-hexosaminidase is an enzyme that coexists with histamine in mast cells, and is used as an index to confirm anti-allergic effects in proportion to the amount of histamine leaked by degranulation. As a result of treating RBL-2H3 cells with the Picalibacterium prausnitzii KBL1027 strain according to an example of the present invention, it was confirmed that the secretion of β-hexosaminidase was inhibited and the anti-allergic effect was excellent.

(2) Cytokine measurement

The cytokine IL-10 is a cytokine secreted by Th2 cells that suppresses inflammation and allergy. RBL-2H3 cells were treated with the Picalibacterium prausnitzii KBL1027 strain according to an example of the present invention, and then IL-10 was measured, confirming that the anti-allergic effect was excellent.

Example 9. F. prausnitzii Evaluation of periodontitis treatment activity of strain KBL1027

As a result of treating the oral caries causative bacterium, Streptococcus mutans, with the Picalibacterium prausnitzii KBL1027 strain according to an embodiment of the present invention, it was confirmed that the oral caries causative bacterium was inhibited and the treatment effect of periodontitis was excellent.

Example 10. F. prausnitzii Antioxidant activity evaluation of strain KBL1027

(One) DCFH-DA assay (measuring absorbance)

As a result of measuring the intracellular ROS scavenging ability, it was confirmed that the P. prausnitzii KBL1027 strain according to an example of the present invention had excellent antioxidant activity.

(2) DPPH assay (measuring absorbance)

As a result of measuring the DPPH radical scavenging ability, it was confirmed that the P. prausnitzii KBL1027 strain according to an example of the present invention had excellent antioxidant activity.

Example 11. Anti-inflammatory efficacy evaluation using LPS animal model

For anti-inflammatory evaluation of KBL1027 strain, LPS (Lipopolysaccharide), an inflammation-inducing factor, and KBL1027 strain were simultaneously administered to the mouse peritoneal cavity, and the amount of cytokine secretion secreted from macrophages during an acute inflammatory reaction was measured. When the KBL1027 strain and LPS were administered simultaneously, the expression of IL-10 cytokine increased, and the expression of inflammatory mediating cytokines IL-6 and TNF-a decreased compared to the LPS-only challenge group. Through this, the anti-inflammatory efficacy of the KBL1027 strain was confirmed in an animal model.

Example 12. Evaluation of psoriasis (autoimmune) treatment activity using an animal model

In order to evaluate the therapeutic efficacy against the KBL1027 strain in a psoriasis animal model among autoimmune diseases, female BALB / c mouse back hair was depilated using a hair removal cream, and imiquimod cream was applied to the mouse back skin every day for 6 days, A psoriasis induction model was constructed. In the psoriasis-induced animal model, the skin tissue of the KBL1027 strain application group was removed and tissue staining was performed with hematoxylin and eosin. It was confirmed that was significantly reduced.

Example 13. Assessment of asthma treatment activity using animal models

Allergic asthma was induced by injecting ovalbumin into the peritoneal cavity of an animal subject to evaluate the therapeutic efficacy against the KBL1027 strain in an allergic respiratory disease animal model. When the inflammatory index was measured in the lung tissue of the asthmatic mouse model mouse and the degree of inflammatory cell infiltration around the bronchi was scored and evaluated numerically, it was confirmed that the pulmonary inflammatory index was significantly reduced in the KBL1027 strain-administered group.

Korea Research Institute of Bioscience and Biotechnology KCTC14230BP 20200707 Korea Research Institute of Bioscience and Biotechnology KCTC14231BP 20200707

<110> KoBioLabs, Inc. <120> Faecalibacterium prausnitzii strain and uses thereof <130> DPP20214836KR <160> 14 <170> KoPatentIn 3.0 <210> 1 <211> 20 <212> DNA <213> artificial sequence <220> <223> (synthetic) 27F primer <220> <221> misc_feature <222> (11) <223> y is t or c <220> <221> misc_feature <222> (12) <223> m is a or c <400> 1 agagtttgat ymtggctcag 20 <210> 2 <211> 21 <212> DNA <213> artificial sequence <220> <223> (synthetic) 1492R primer <220> <221> misc_feature <222> (6) <223> y is t or c <400> 2 tacggytacc ttgttacgac t 21 <210> 3 <211> 1383 <212> DNA <213> Faecalibacterium prausnitzii <400> 3 gacgaacgct ggcggcgcgc ctaacacatg caagtcgaac gagtgagaga gagcttgctt 60 120 aacagttgga aacgactgct aataccgcat aagcccacga cccggcatcg ggtagaggga 180 aaaggagcaa tccgctttga gatggcctcg cgtccgatta gctagttggt gaggtaactg 240 gcccaccaag gcgacgatcg gtagccggac tgagaggttg aacggccaca ttgggactga 300 gacacggccc agactcctac gggaggcagc agtggggaat attgcacaat gggggaaacc 360 ctgatgcagc gacgccgcgt ggaggaagaa 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ttcccggtgt agcggtggaa 660 tgcgtagata tcgggaggaa caccagtggc gaaggcggcc tactgggcac caactgacgc 720 tgaggctcga aagtgtgggt agcaaacagg attagatacc ctggtagtcc acactgtaaa 780 cgatgattac taggggttgg aggattgacc ccttcagtgc cgcagttaac acaataagta 840 atccacctgg ggagtacgac cgcaaggttg aaactcaaag gaattgacgg gggcccgcac 900 aagcagtgga gtatgtggtt taattcgacg caacgcgaag aaccttacca agtcttgaca 960 tcctgcgacg cacatagaaa tatgtgtttc cttcgggacg cagagacagg tggtgcatgg 1020 ttgtcgtcag ctcgtgtcgt gagatgttgg gttaagtccc gcaacgagcg caacccttat 1080 ggtcagttac tacgcaagag gactctggcc agactgccgt tgacaaaacg gaggaaggtg 1140 gggatgacgt caaatcatca tgccctttat gacttgggct acacacgtac tacaatggcg 1200 ttaaacaaag agaagcaaga ccgcgaggtg gagcaaaact cagaaacaac gtcccagttc 1260 ggactgcagg ctgcaactcg cctgcacgaa gtcggaattg ctagtaatcg cagatcagca 1320 tgctgcggtg aatacgttcc cgggccttgt acacaccgcc cgtcacacca tgagagccgg 1380 ggggacccga agtcggtagt ctaaccgcaa ggag 1414 <210> 5 <211> 59 <212> DNA <213> artificial sequence <220> <223> (synthetic) 515F primer <220> <221> misc_feature <222> (49) <223> m is a or c <400> 5 atgatacggc gaccaccgag atctacacta tggtaattgt gtgccagcmg ccgcggtaa 59 <210> 6 <211> 56 <212> DNA <213> artificial sequence <220> <223> (synthetic) 806R primer <220> <221> misc_feature <222> (44) <223> h is a, c, or t <220> <221> misc_feature <222> (45) <223> v is g, c, or a <220> <221> misc_feature <222> (50) <223> w is a or t <400> 6 caagcagaag acggcatacg agatagtcag tcagccggac tachvgggtw tctaat 56 <210> 7 <211> 21 <212> DNA <213> artificial sequence <220> <223> (synthetic) HPRT primer FWD <400> 7 ttatggacag gactgaaaga c 21 <210> 8 <211> 22 <212> DNA <213> artificial sequence <220> <223> (synthetic) HPRT primer RVS <400> 8 gctttaatgt aatccagcag gt 22 <210> 9 <211> 20 <212> DNA <213> artificial sequence <220> <223> (synthetic) IL-10 primer FWD <400> 9 ataactgcac ccacttccca 20 <210> 10 <211> 20 <212> DNA <213> artificial sequence <220> <223> (synthetic) IL-10 primer RVS <400> 10 tcatttccga taaggcttgg 20 <210> 11 <211> 24 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Claims (23)

Faecalibacterium prausnitzii KBL1027 strain having accession number KCTC14231BP. According to claim 1, wherein the strain is characterized in that it has a 16S rRNA sequence represented by SEQ ID NO: 4, the strain. The strain according to claim 1, wherein the strain produces extracellular polysaccharide. The strain according to claim 1, wherein the strain contains genes related to cell wall and capsule functions. The strain according to claim 4, wherein the genes related to the cell wall and capsule function perform the function of exopolysaccharide biosynthesis or rhamnose containing glycans. The strain according to claim 4, wherein the cell wall and capsule function-related genes include one or more genes selected from the group consisting of the following (1) to (7):
(1) Exopolysaccharide biosynthesis glycosyltransferase;
(2) Tyrosine-protein kinase;
(3) Alpha-D-GlcNAc alpha-1,2-L-rhamnosyltransferase;
(4) Alpha-L-Rha alpha-1,3-L-rhamnosyltransferase;
(5) capsular polysaccharide biosynthesis protein,
(6) Lipoprotein releasing system ATP-binding protein;
(7) Putative N-acetylgalactosaminyl-diphosphoundecaprenol glucuronosyltransferase. The strain according to claim 1, wherein the strain comprises one or more genes selected from the group consisting of the following (1) to (13):
(1) Exopolysaccharide biosynthesis glycosyltransferase;
(2) Tyrosine-protein kinase;
(3) Alpha-D-GlcNAc alpha-1,2-L-rhamnosyltransferase;
(4) Alpha-L-Rha alpha-1,3-L-rhamnosyltransferase;
(5) capsular polysaccharide biosynthesis protein,
(6) Lipoprotein releasing system ATP-binding protein;
(7) Putative N-acetylgalactosaminyl-diphosphoundecaprenol glucuronosyltransferase;
(8) Predicted transcriptional regulator of N-Acetylglucosamine utilization, GntR family,
(9) Membrane-bound lytic murein transglycosylase B;
(10) Inosine-uridine preferring nucleoside hydrolase;
(11) LysR family transcriptional regulator,
(12) Cation efflux system protein;
(13) Cobalt-zinc-cadmium resistance protein. The strain according to claim 1, wherein the strain has a characteristic of inducing anti-inflammatory cytokine production. The strain according to claim 1, wherein the strain has a characteristic of enhancing tight junctions between intestinal epithelial cells. The strain according to claim 1, wherein the strain has a characteristic of improving the balance of microorganisms in the intestine. The strain according to claim 10, wherein the intestinal microbial balance is improved by one or more selected from the group consisting of the following (1) to (5):
(1) Inducing an increase in the diversity of flora in the intestines of animals;
(2) improving the imbalance of flora in the intestines of animals;
(3) induces an increase in the relative abundance of strains of the genus Prevotella and/or the family Paraprevotellaceae in the intestinal flora of animals;
(4) preventing an increase in the relative abundance of harmful bacteria having lipopolysaccharide cell walls in the intestinal flora of animals;
(5) preventing an increase in the relative abundance of strains of the genus Bacteroides and/or the phylum Proteobacteria in the animal's intestinal flora; The strain according to claim 1, wherein the strain has one or more characteristics selected from the group consisting of the following (1) to (2):
(1) preventing weight loss due to inflammatory diseases;
(2) preventing intestinal length reduction caused by inflammatory diseases; A culture of the strain according to any one of claims 1 to 12, a lysate of the strain, an extract of the strain, an extract of the culture, an extract of the lysate, or an extracellular polysaccharide produced by the strain. The strain according to any one of claims 1 to 12, the culture of the strain, the lysate of the strain, the extract of the strain, the extract of the culture, the extract of the lysate, and the extracellular polysaccharide produced by the strain A composition for the prevention, improvement, or treatment of inflammatory diseases comprising at least one selected from the group consisting of. The method of claim 14, wherein the inflammatory disease is an immune disease, inflammatory bowel disease, inflammatory skin disease, inflammatory collagen vascular disease, atopic dermatitis, urticaria, allergic disease, autoimmune disease, autoimmune inflammatory disease, autoimmune skin disease, autoimmunity Inflammatory skin disease, eczema, asthma, allergic asthma, bronchial asthma, rhinitis, allergic rhinitis, conjunctivitis, allergic conjunctivitis, food allergy, anaphylactic shock, rheumatoid arthritis, rheumatoid fever, lupus, systemic Lupus erythematosus, systemic scleroderma, psoriasis, psoriatic arthritis, asthma, Guilian-Barre syndrome, myasthenia gravis, dermatomyositis, polymyositis, multiple sclerosis, type 1 diabetes, autoimmune encephalomyelitis, polyarteritis nodosa, Hashimoto Thyroiditis, systemic sclerosis, temporal arteritis, childhood diabetes, alopecia areata, pemphigus, aphthous stomatitis, autoimmune hemolytic anemia, Wegener's granulomatosis, Sjogren's syndrome, Addison's disease, Behçet's disease, edema, conjunctivitis, periodontitis, rhinitis, otitis media, chronic sinusitis, Sore throat, tonsillitis, bronchitis, pneumonia, gastric ulcer, gastritis, colitis, gout, eczema, acne, contact dermatitis, seborrheic dermatitis, ankylosing myelitis, fibromyalgia, osteoarthritis, periarthritis, tendinitis, At least one member selected from the group consisting of tenosynovitis, myositis, hepatitis, cystitis, nephritis, sepsis, vasculitis, and bursitis, a composition. 16. The method of claim 15, wherein the inflammatory bowel disease is inflammatory bowel disease, ulcerative colitis (UC), enteritis, irritable bowel syndrome (IBS), and Crohn's disease (CD) One or more selected from the group consisting of, the composition. The composition according to claim 14, wherein the prevention, improvement, or treatment of the inflammatory disease is caused by the extracellular polysaccharide of the strain. The strain according to any one of claims 1 to 12, the culture of the strain, the lysate of the strain, the extract of the strain, the extract of the culture, the extract of the lysate, and the extracellular polysaccharide produced by the strain A food composition for preventing or improving inflammatory diseases, comprising at least one selected from the group consisting of. The strain according to any one of claims 1 to 12, the culture of the strain, the lysate of the strain, the extract of the strain, the extract of the culture, the extract of the lysate, and the extracellular polysaccharide produced by the strain Probiotics composition for preventing or improving inflammatory diseases, comprising at least one selected from the group consisting of. Faecalibacterium prausnitzii strain according to any one of claims 1 to 12, culture of the strain, lysate of the strain, extract of the strain, extract of the culture, lysate of the lysate A composition for improving intestinal microbial balance, comprising at least one selected from the group consisting of extracts and extracellular polysaccharides produced by the strain. Faecalibacterium prausnitzii strain having anti-inflammatory activity, a culture of the strain, a lysate of the strain, an extract of the strain, an extract of the culture, an extract of the lysate, or produced by the strain As an extracellular polysaccharide,
The strain is a Faecalibacterium prausnitzii KBL1027 strain having accession number KCTC14231BP, a strain, a culture of the strain, a lysate of the strain, an extract of the strain, an extract of the culture, an extract of the lysate , or an extracellular polysaccharide produced by the strain. Faecalibacterium prausnitzii strain, a culture of the strain, a lysate of the strain, an extract of the strain, an extract of the culture, an extract of the lysate, or the above As an extracellular polysaccharide produced by the strain,
The strain is a Faecalibacterium prausnitzii KBL1027 strain having accession number KCTC14231BP, a strain, a culture of the strain, a lysate of the strain, an extract of the strain, an extract of the culture, an extract of the lysate , or an extracellular polysaccharide produced by the strain. The method of claim 22, wherein the substance having anti-inflammatory activity is an extracellular polysaccharide, a strain, a culture of the strain, a lysate of the strain, an extract of the strain, an extract of the culture, an extract of the lysate, or the strain Extracellular polysaccharides produced by

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