Bifidobacterium mixed strain having excellent Anti-inflammatory activity and productivity, and use thereof

Abstract

The present invention relates to a mixed strain having excellent anti-inflammatory activity and productivity, comprising Bifidobacterium animalis subsp. lactis LM1017 (accession number KCCM12629P), Bifidobacterium breve LM1092 (accession number KCCM13464P), Bifidobacterium longum LM1024 (accession number KCCM12919P), and Bifidobacterium bifidum LM1108 (accession number KCCM13463P).

Description

Bifidobacterium mixed strain with excellent anti-inflammatory activity and productivity and its use

[0001] The present invention relates to a mixed bifidobacterium strain with excellent productivity and anti-inflammatory activity and its use.

[0002] Lactic acid bacteria have been a constant companion to humans throughout history, and their utility as microorganisms with profound health benefits is steadily increasing. Recently, active research on lactic acid bacteria has led to their expansion into not only general food but also health foods and pharmaceuticals, expanding their application. These lactic acid bacteria reside in the intestines of animals, breaking down ingested nutrients and fiber to use them as energy sources. They also produce lactic acid and antibiotics, inhibiting the growth of harmful intestinal bacteria and playing a crucial role in maintaining intestinal health. Furthermore, lactic acid bacteria are used to promote animal growth and improve feed utilization, increase resistance to disease, suppress the growth of harmful bacteria, reduce mortality, suppress the production of putrefactive toxins, and produce various vitamins. However, to achieve these benefits, lactic acid bacteria must reach the intestines without hindrance and function effectively. This requires minimal destruction by gastric acid when administered orally and strong resistance to bile acids.

[0003] Meanwhile, inflammation occurs when an organism's immune system attempts to respond to harmful substances, such as invading pathogens. Immunoregulatory mechanisms, including immunoregulatory cells, cytokines, and apoptosis, regulate the immune response, which occurs as an excessive response to protect against pathogens. Deficiency in these mechanisms can lead to various inflammatory diseases, including IBD. Cytokine production triggered by bacterial components such as lipopolysaccharides (LPS), lipoteichoic acid (LTA), and peptidoglycan can lead to systemic inflammatory response syndrome (SIRS). LPS activates cells of the innate immune system to produce various inflammatory cytokines, such as interleukin-1 (IL-1), IL-6, IL-8, and tumor necrosis factor-α (TNF-α), as well as nitric oxide (NO). NO is a highly reactive molecule synthesized from L-arginine by the NO synthase (NOS) isoenzyme family. The isoenzyme present in macrophages is an inducible form (iNOS), which can produce high concentrations of NO in many cells upon stimulation by bacterial endotoxins, LPS, or other proinflammatory cytokines, including TNF-α, IFN-γ, IL-1, and IL-6. Among several inflammatory diseases, inflammatory bowel disease (IBD) is closely associated with probiotics. IBD patients may experience abdominal pain, diarrhea, intestinal obstruction, and / or bloody stool. To date, anti-inflammatory drugs, steroids, and anti-TNF agents have been shown to effectively relieve symptoms, but a fundamental medical treatment for IBD is still unknown.

[0004] The present inventors mixed four strains belonging to the genus Bifidobacterium by adding the Bifidobacterium breve LM1092 strain, which, like the LM1017 strain, has low anti-inflammatory activity as a single strain but high productivity, and the Bifidobacterium breve LM1108 strain, which, like the LM1024 strain, has high anti-inflammatory activity as a single strain but low productivity, to the LM1017 and LM1024 strains. Through this, the present inventors intend to provide a mixed strain with excellent anti-inflammatory activity and productivity and its use.

[0005] In order to solve the above problem, the present invention provides a mixed strain having excellent anti-inflammatory activity and productivity, wherein the mixed strain includes Bifidobacterium animalis lactis LM1017 (Accession No. KCCM12629P), Bifidobacterium breve LM1092 (Accession No. KCCM13464P), Bifidobacterium longum LM1024 (Accession No. KCCM12919P), and Bifidobacterium breve LM1108 (Accession No. KCCM13463P).

[0006] The mixed strain of this invention can be used as a probiotic, and specifically, any one or more of the mixed strain, culture of the strain, lysate of the strain, and extract of the strain can be included as an effective ingredient in an anti-inflammatory composition, a food composition, a health functional food composition, or a pharmaceutical composition.

[0007] Another aspect of the present invention provides a method for improving anti-inflammatory activity, comprising administering to a subject in need thereof at least one of a mixed strain, a culture of the mixed strain, a lysate of the mixed strain, and an extract of the mixed strain.

[0008] Another aspect of the present invention provides a use for improving anti-inflammatory activity and productivity of at least one of the present invention's mixed strains, mixed strain cultures, mixed strain lysates, and mixed strain extracts.

[0009] Another aspect of the present invention provides a use of any one or more of the present invention's mixed strain, mixed strain culture, mixed strain lysate, and mixed strain extract for producing an anti-inflammatory drug.

[0010] The effects of the present invention include not only high anti-inflammatory activity but also high productivity, resulting in excellent probiotic business potential.

[0011] Figure 1 shows the results of a productivity experiment according to Example 1.

[0012] Figure 2 shows the results of an anti-inflammatory activity experiment according to Example 2.

[0013] Below, with reference to the attached drawings, embodiments of the present invention are described in detail to facilitate easy implementation by those skilled in the art. However, the present invention can be implemented in various different forms and is not limited to the embodiments described herein. In the drawings, irrelevant parts have been omitted for clarity, and similar reference numerals have been used throughout the specification to indicate similar elements.

[0014] Throughout this specification, whenever a part is said to 'include' a component, this does not mean that it excludes other components, but rather that it may include other components, unless otherwise specifically stated.

[0015] Throughout this specification, the terms “step of” or “step of” do not mean “step for”.

[0016] Throughout this specification, the term 'combination(s) thereof' included in the expressions in the Makushi format means one or more mixtures or combinations selected from the group consisting of the components described in the expressions in the Makushi format, and means including one or more selected from the group consisting of said components.

[0017] Throughout this specification, references to 'A and / or B' mean 'A or B, or A and B.'

[0018] Throughout this specification, 'Bifidobacterium animalis lactis LM1017' refers to the Bifidobacterium animalis subsp. lactis LM1017 strain. It may also be indicated as LM1017, LM1017 strain, etc.

[0019] Throughout this specification, 'Bifidobacterium breve LM1092' refers to the Bifidobacterium breveLM1092 strain. It may also be indicated as LM1092, LM1092 strain, etc.

[0020] Throughout this specification, 'Bifidobacterium longum LM1024' refers to the Bifidobacterium longum LM1024 strain. It may also be indicated as LM1024, LM1024 strain, etc.

[0021] Throughout this specification, 'Bifidobacterium bifidum LM1108' refers to the Bifidobacterium bifidum LM1108 strain. It may also be indicated as LM1108, LM1108 strain, etc.

[0022] The above strains may be interpreted to include the strain itself, a culture of the strain, a lysate of the strain, an extract of the strain, and a cytoplasmic fraction obtained by lysing the strain.

[0023] In one specific example, the strain may be a strain that is a mutant of a naturally occurring strain.

[0024] In one specific example, the strain may be a mixed strain with excellent anti-inflammatory activity and productivity. Furthermore, the anti-inflammatory activity may include inhibition of nitric oxide production.

[0025] Throughout the present specification, the term "culture" refers to the entire medium including the strain itself, an extract of the strain, a metabolite of the strain, or extra nutrients obtained by culturing the strain in a medium supplied with nutrients for a certain period of time, and also includes a culture solution from which the strain has been removed after culturing the strain. In addition, it may include a concentrate of the entire medium or culture solution, and a dried product of the concentrate. Specifically, the culture of the present invention may use a medium easily selected by a person skilled in the art according to the purpose among media used for culturing microorganisms, for example, MRS (de MAN, ROGOSA and SHARPE) or BL medium may be used, but is not limited thereto as long as the strain of the present invention can be cultured.

[0026] Throughout this specification, the term "lysate" may be used interchangeably with "lysate," which refers to a solution or suspension of cells of a microorganism, such as a broken strain, in an aqueous medium. A cell lysate includes, for example, macromolecules such as DNA, RNA, proteins, peptides, carbohydrates, lipids, etc., and / or micromolecules such as amino acids, sugars, fatty acids, etc., or fractions thereof. The lysate also includes cell debris, which may be smooth or granular in structure.

[0027] Throughout this specification, the term "extract" refers to an isolate separated from a culture medium of a strain. The extract may also include a fraction or culture filtrate.

[0028] Generally, strains used as probiotics are expected to have the same or similar effects, whether they are the strain itself, strain culture, strain lysate, or strain extract.

[0029] Throughout this specification, "productivity" refers to the productivity of a strain, which may be related to the number of strains that can be obtained after cultivation. For example, high strain productivity may mean a large number of strains obtained after cultivation. In the probiotics industry, the productivity of lactic acid bacteria directly translates to the productivity of probiotic products, so strain productivity is directly linked to the business viability of probiotics. Therefore, it is important to use strains that not only have good functions but also have high productivity.

[0030] In addition, throughout the specification of the present invention, 'excellent productivity' means that it is advantageous in obtaining the amount of lactic acid bacteria required as a probiotic product, and does not mean that when the strains according to the present invention are cultured in a mixture, the productivity increases compared to when they are cultured as a single strain. In other words, the excellent productivity of the present invention means that, in securing the amount of lactic acid bacteria required as a probiotic product, (1) the required amount can be more easily secured when a low-productivity strain and a high-productivity strain are used together compared to when only a low-productivity strain is used; it does not mean that the culture is better when the mixed strain is cultured in a single medium.

[0031] Throughout this specification, the term 'logarithmic phase' refers to the period in a bacterial growth curve where the population of microorganisms doubles.

[0032] Throughout this specification, the term 'stationary phase' refers to the period in a bacterial growth curve when a population of microorganisms stops growing and the growth curve becomes horizontal.

[0033] Throughout this specification, 'food' specifically includes meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages, vitamin complexes, health functional foods and health foods, and may include all conventional foods such as natural foods and processed foods.

[0034] The food of the present invention can be manufactured using methods commonly used in the art, and during the manufacturing process, raw materials and ingredients commonly added in the art can be added. In addition, the food formulation can be manufactured without limitation as long as it is a formulation recognized as a food. The food composition of the present invention can be manufactured in various forms, and unlike general drugs, it has the advantage of not having side effects that can occur with long-term use of drugs because it uses food as a raw material. In addition, it is highly portable, so it can be consumed as a supplement.

[0035] The above food composition may additionally include a physiologically acceptable carrier. The type of the carrier is not particularly limited, and any carrier commonly used in the technical field of the present invention may be used.

[0036] Additionally, the food composition may include additional ingredients commonly used in food compositions to improve odor, taste, sight, etc. For example, it may include vitamins A, C, D, E, B1, B2, B6, B12, K1, K2, niacin, biotin, folate, pantothenic acid, etc. In addition, it may include minerals such as zinc (Zn), iron (Fe), calcium (Ca), chromium (Cr), magnesium (Mg), manganese (Mn), copper (Cu), etc.

[0037] As an example, the food composition of the present invention can be used as a health beverage composition, and in this case, various natural carbohydrates or sweeteners, etc. can be contained as additional ingredients like regular beverages. The above natural carbohydrates can be monosaccharides, disaccharides, polysaccharides, sugar alcohols, etc. The sweetener can be a natural sweetener such as Thaumatin, Stevia extract, Xylitol, allulose, inulin, etc.; or a synthetic sweetener such as Saccharin, Aspartame, Sucralose, etc.

[0038] In addition to the above, the health beverage composition may contain various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid, salts of pectic acid, alginic acid, salts of alginic acid, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, or carbonating agents. Furthermore, the composition may contain fruit pulp for the production of natural fruit juice, fruit juice beverages, or vegetable beverages. These ingredients may be used independently or in combination.

[0039] Throughout this specification, 'health functional food' may refer to food manufactured and processed using raw materials or ingredients with functionality useful to the human body according to the Health Functional Food Act ('HEALTH FUNCTIONAL FOODS ACT' of Korea No. 6727), and 'functionality' may refer to obtaining a useful effect for health purposes such as regulating nutrients for the structure and function of the human body or physiological action.

[0040] The above health functional food refers to a food that has a more active health maintenance or promotion effect than general food, and health supplement food refers to a food for the purpose of health supplementation. In some cases, the terms health functional food, health food, and health supplement may be used interchangeably. Specifically, the above health functional food refers to a food that is prepared by adding the present invention to food materials such as beverages, tea, spices, gum, and confectionery, or by manufacturing it in the form of capsules, powder, suspension, etc., and may mean that when consumed, it brings about a specific health effect. However, unlike general drugs, it has the advantage of not having side effects that may occur with long-term use of drugs because it uses food as a raw material.

[0041] Throughout the present specification, the term 'pharmaceutical composition' may be used in the form of oral medications such as powder, granule, tablet, capsule, suspension, emulsion, syrup, aerosol, ointment, suppository or sterile injection according to a conventional method, but is not limited thereto.

[0042] A pharmaceutical composition according to one embodiment of the present invention may be a 'pharmaceutical composition' or a 'quasi-drug composition'.

[0043] The term 'quasi-drug' used throughout this specification may refer to products that have a milder effect than pharmaceutical products among products used for the purpose of diagnosing, treating, improving, alleviating, managing or preventing diseases in humans or animals. For example, according to the Pharmaceutical Affairs Act, quasi-drugs are products excluding products used for the purpose of pharmaceutical products, and may include products used for treating or preventing diseases in humans or animals, products that have a mild effect on the human body or do not act directly.

[0044] In one embodiment of the present invention, the pharmaceutical composition may be administered in a pharmaceutically effective amount, wherein the 'pharmaceutically effective amount' means an amount sufficient to treat or prevent a disease at a reasonable benefit-to-risk ratio applicable to medical treatment or prevention. The level of the effective amount may be determined according to factors including the severity of the disease, the activity of the drug, the patient's age, weight, health, sex, the patient's sensitivity to the drug, the time of administration of the composition of the present invention used, the route of administration and the excretion rate, the treatment period, drugs used in combination or simultaneously with the composition of the present invention used, and other factors well known in the medical field. The pharmaceutical composition of the present invention may be administered alone or in combination with a component known to exhibit a therapeutic effect on a known intestinal disease. It is important to take all of the above factors into consideration and administer an amount that can achieve the maximum effect with the minimum amount without side effects.

[0045] The pharmaceutical composition of the present invention is not particularly limited thereto, but may be administered via routes such as intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, intradermal administration, transdermal patch administration, oral administration, intranasal administration, intrapulmonary administration, and rectal administration, depending on the intended purpose. However, when administered orally, it may be administered in an unformulated form, and since the present invention may be denatured or decomposed by gastric acid, the oral composition may be administered orally in a form that coats the active agent or is formulated to protect it from decomposition in the stomach, or in the form of an oral patch. In addition, the composition may be administered by any device that allows the active agent to move to the target cell.

[0046] Hereinafter, implementation examples and embodiments of the present invention will be described in detail with reference to the attached drawings. However, the present invention may not be limited to these implementation examples and embodiments and drawings.

[0047] The present invention provides a mixed strain having excellent anti-inflammatory activity and productivity, wherein the mixed strain includes Bifidobacterium animalis lactis LM1017 (Accession No. KCCM12629P), Bifidobacterium breve LM1092 (Accession No. KCCM13464P), Bifidobacterium longum LM1024 (Accession No. KCCM12919P), and Bifidobacterium breve LM1108 (Accession No. KCCM13463P).

[0048] In order to discover a mixed strain with both excellent anti-inflammatory properties and productivity, the present inventors mixed the LM1017 and LM1024 strains, which are known to be anti-inflammatory, with the LM1092 strain and the LM1108 strain.

[0049] In one specific example of the present invention, the mixing ratio of each strain (LM1017 and LM1092 [group A]: LM1024 and LM1108 [group B]) may preferably be 32:1 to 8:1, 32:1 to 1:1, 32:1 to 1:8, 32:1 to 1:32, 8:1 to 1:1, 8:1 to 1:8, 8:1 to 1:32, 1:1 to 1:8, 1:1 to 1:32, 1:8 to 1:32, but is not limited thereto.

[0050] In one specific example of the present invention, the anti-inflammatory activity may include inhibition of nitric oxide production.

[0051] And the present invention provides an anti-inflammatory composition, an anti-inflammatory food composition, an anti-inflammatory health functional food composition, and an anti-inflammatory pharmaceutical composition, which comprises at least one of a mixed strain, a culture of a mixed strain, a lysate of a mixed strain, and an extract of a mixed strain as an active ingredient.

[0052] In one specific example of the present invention, the characteristics of the mixed strains included in the above compositions may be the same as the characteristics of the mixed strains described above.

[0053] Another aspect of the present invention provides an anti-inflammatory improvement method comprising a step of administering to a subject in need thereof at least one of a mixed strain, a culture of the mixed strain, a lysate of the mixed strain, and an extract of the mixed strain.

[0054] Another aspect of the present invention provides the use of improving anti-inflammatory properties and productivity by at least one of the present invention's mixed strains, cultures of mixed strains, lysates of mixed strains, and extracts of mixed strains.

[0055] Another aspect of the present invention provides a use of at least one of the mixed strain, the culture of the mixed strain, the lysate of the mixed strain, and the extract of the mixed strain for producing a drug for improving anti-inflammatory activity.

[0056] As a result of confirming the anti-inflammatory activity and productivity of each strain used in the mixed strain of this invention, the LM1017 and LM1092 strains had low anti-inflammatory activity as individual strains but high productivity, and the LM1024 and LM1108 strains had high anti-inflammatory activity as individual strains but low productivity. However, by mixing all of the above strains, the low productivity of the LM1024 and LM1108 strains was supplemented, and as a result of confirming the anti-inflammatory activity of the mixed strain, it was confirmed that the anti-inflammatory activity was also excellent, and based on this, the present invention was invented.

[0057] Example 1. Productivity verification experiment

[0058] Experiments were conducted to determine the productivity of Bifidobacterium animalis lactis LM1017, Bifidobacterium breve LM1092, Bifidobacterium longum LM1024, and Bifidobacterium bifidum LM1108 strains.

[0059] Each strain was revived once (37°C, 18 hours) in BL liquid medium. Afterwards, 1% of the bacterial cells were inoculated into the same medium and cultured at 37°C. The productivity of the bacterial cells was measured by measuring the optical density (OD) of the culture solution using a turbidity measurement method after 0, 2, 4, 6, 8, 10, 12, and 24 hours of culture. 600nm It was judged based on .

[0060] As a result of the experiment, in the logarithmic and stationary phases, the OD of Bifidobacterium bifidum LM1108, Bifidobacterium longum LM1024, Bifidobacterium breve LM1092, and Bifidobacterium animalis lactis LM1017 were in that order. 600nm It was confirmed that the increase was significant (Fig. 1, Table 1).

[0061] Therefore, it was confirmed that the productivity of the LM1017 and LM1092 strains was approximately twice as high as that of the LM1024 and LM1108 strains (based on a culture time (h) = 6). However, the inventors of the present invention compensated for the low productivity of the LM1024 and LM1108 strains by mixing the LM1017 and LM1092 strains.

[0062] Incubation time (h)OD 600nm LM1017LM1092LM1024LM110800.00500.00700.00300.001020.01970.02690.01620.012040.03140.04520.02900.027060.17360.15950 .12320.062180.29800.22400.16500.1258100.32530.23860.20430.1563120.32420.24430.19740.1545240.33670.25040.20890.1541

[0063] Example 2. Anti-inflammatory activity confirmation experiment

[0064] An experiment was conducted to determine the anti-inflammatory activity of a mixed strain containing Bifidobacterium animalis lactis LM1017, Bifidobacterium breve LM1092, Bifidobacterium longum LM1024, and Bifidobacterium breve LM1092. Anti-inflammatory activity was confirmed by inhibition of nitric oxide production.

[0065] Specifically, DMEM medium (Welgene, Korea) containing 10% fetal bovine serum (Welgene, Korea) and penicillin-streptomycin (Welgene, Korea) was used to culture RAW 264.7 cells, a mouse-derived macrophage cell line. RAW 264.7 cells were seeded in a 96-well plate at a density of 3X10 4After dispensing at a concentration of cells / well, the wells were cultured in a CO2 incubator at 37℃ for O / N. After removing the medium from each well, Bifidobacterium animalis lactis LM1017, Bifidobacterium breve LM1092, Bifidobacterium longum LM1024, and Bifidobacterium breve LM1092 were diluted in the medium and treated alone or in combination for 2 hours. To induce inflammation in RAW 264.7 cells, LPS was treated to each well and cultured for an additional 24 hours in a CO2 incubator at 37℃. The Griess Reagent system from Promega was used to measure the production of nitric oxide. 50 μl of the cultured medium was transferred to a new 96-well plate. At this time, the nitric oxide standard solution was diluted to 1.56–100 μM for a nitric oxide standard curve. After adding 50 μl of sulfanilamide solution to each well, the mixture was incubated at room temperature for 5 minutes, protected from light. An additional 50 μl of NED solution was added to each well, and the mixture was incubated at room temperature for 5 minutes, protected from light. The absorbance was measured at 540 nm using a microplate reader, and the amount of nitric oxide produced was calculated using the nitric oxide standard curve.

[0066] First, as a result of confirming the nitric oxide production inhibition ability of each strain, Bifidobacterium animalis lactis LM1017 and Bifidobacterium breve LM1092 showed about 5% nitric oxide production inhibition ability against nitric oxide induced by inflammation caused by LPS, LM1024 showed about 11% nitric oxide production inhibition ability, and LM1108 showed about 16% nitric oxide production inhibition ability (see Fig. 2, Table 2; left). In other words, it was confirmed that the anti-inflammatory activity of the LM1017 and LM1092 strains was low, whereas the anti-inflammatory activity of the LM1024 and LM1108 strains was high.

[0067] Accordingly, the inventors of the present invention mixed all four strains above to invent a probiotic with high productivity. Then, they divided the strains into groups: LM1017 and LM1092 strains with high productivity only (group A), and LM1024 and LM1108 strains with high anti-inflammatory activity only (group B). In order to confirm the effect of each group on the anti-inflammatory activity of the mixed strain, the mixed strains were prepared by varying the mixing ratio of each group, and the nitric oxide production inhibition ability of each mixed strain was confirmed (see Fig. 2, Table 2; right A:B). The horizontal line included in Fig. 2 is the 90% nitric oxide production inhibition ability baseline, which represents the anti-inflammatory activity standard required for commercialization.

[0068] As a result, regardless of the mixing ratio of A:B, it showed at least 10% or more of nitric oxide production inhibition ability in all cases, and compared to the theoretical value, the experimental value showed a value that was about 5-7% lower. In other words, the synergistic effect of the four strains according to the present invention on NO production inhibition was confirmed. In particular, the mixed strain according to the present invention showed a superior nitric oxide production inhibition ability than the single strains LM1017 and LM1092 with high productivity in all mixing ratios. This means that the mixed strain according to the present invention overcomes the disadvantage of low anti-inflammatory activity or productivity when each strain is used alone, and is excellent in both anti-inflammatory activity and productivity.

[0069] Nitric oxide (%) theoretical value 1) Experimental value / Theoretical value Negative control group 0.00--LPS treatment group (positive control group) 100.00--Single treatment group LM 101795.04--LM 109294.38--LM 102489.45--LM 110884.40--A:B 32:189.69 94.800.95 8:188.68 94.27 0.94 1:186.63 91.700.94 1:884.65 89.140.95 1:3282.1688.610.93

[0070] 1) Theoretical value: Calculated according to the mixing ratio based on the NO value when treated alone.

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Claims

1. In a mixed strain with excellent anti-inflammatory activity and productivity, The above mixed strain comprises Bifidobacterium animalis lactis LM1017 (accession number KCCM12629P), Bifidobacterium breve LM1092 (accession number KCCM13464P), Bifidobacterium longum LM1024 (accession number KCCM12919P), and Bifidobacterium breve LM1108 (accession number KCCM13463P).

2. In paragraph 1, A mixed strain, wherein the mixing ratio of the above mixed strains is 32:1 to 1:32 based on 'LM1017 and LM1092': 'LM1024 and LM1108'.

3. In paragraph 1, A mixed strain wherein the above anti-inflammatory activity includes inhibition of nitric oxide production.

4. An anti-inflammatory composition comprising at least one of the mixed strain, culture of the mixed strain, lysate of the mixed strain, and extract of the mixed strain according to paragraph 1 as an active ingredient.

5. An anti-inflammatory food composition comprising at least one of the mixed strain, culture of the mixed strain, lysate of the mixed strain, and extract of the mixed strain according to Article 1 as an effective ingredient.

6. An anti-inflammatory health functional food composition comprising at least one of the mixed strain, culture of the mixed strain, lysate of the mixed strain, and extract of the mixed strain according to Article 1 as an active ingredient.

7. An anti-inflammatory pharmaceutical composition comprising at least one of the mixed strain, culture of the mixed strain, lysate of the mixed strain, and extract of the mixed strain according to Article 1 as an active ingredient.

8. A method for improving anti-inflammatory activity, comprising a step of administering to an individual in need thereof at least one of the mixed strain, the culture of the mixed strain, the lysate of the mixed strain, and the extract of the mixed strain according to paragraph 1.

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