Lactococcus lactis strain having anti-inflammatory, anti-oxidant, anti-cancer, anti-microbial and anti-obesity activity and composition comprising the same

Abstract

Each of the Lactococcus lactis strains SNU09LL and SNU10LL of the present invention has excellent anti-inflammatory, antioxidant, anticancer, antibacterial, and anti-obesity activities, and is a safe strain without hemolytic activity or bile salt degradation ability, so it can be utilized in compositions for the prevention, improvement, or treatment of cancer, compositions for anti-inflammation, probiotic or postbiotic preparations, antibacterial compositions, anti-obesity compositions, etc., and can be very useful in related industrial fields.

Description

Technology Field

[0001] The present invention relates to Lactococcus lactis having anti-inflammatory activity, antioxidant activity, anticancer activity, antibacterial activity, and anti-obesity activity ( Lactococcus lactis This relates to strains and their uses. Background Technology

[0002] Postbiotics encompass both the beneficial metabolites produced by probiotics during the fermentation process and the components of microorganisms. Postbiotics are attracting attention as a new alternative material capable of overcoming the limitations of safety, functionality, and stability inherent in conventional probiotics. The domestic market for probiotics as health functional foods for humans is approaching 1 trillion won, and the market for various functional probiotics, extending beyond gut health, is also growing rapidly.

[0003] In particular, Lactococcus lactis ( Lactococcus lactis Lactobacillus lactis is a lactic acid bacterium traditionally widely used in the production of cheese and fermented milk, and is classified as a GRAS (Generally Recognized As Safe) strain with high safety. Recently, Lactococcus lactis has been attracting attention as a new functional microbial material as research results have shown that it possesses not only fermentation functions but also physiological activities such as the production of antimicrobial substances, immune regulation, and inhibition of inflammatory responses.

[0004] However, the biological activities of previously reported Lactococcus lactis strains are often limited to specific functions, so research on strains possessing multiple biological activities simultaneously is relatively lacking. Therefore, there is an increasing demand for the development of technologies to discover novel Lactococcus lactis strains that exhibit various biological activities simultaneously and to utilize them for applications such as functional foods, health functional foods, pharmaceuticals, and animal feed additives.

[0005] Meanwhile, Korean Patent No. 2645456 discloses 'a Lactococcus lactis MG5474 strain having immune-enhancing and gut health-promoting activities and a composition containing the same,' and Korean Patent No. 1841021 discloses 'a novel Lactococcus lactis strain having advanced glycation end product reduction activity and the use thereof,' but there is no description of the 'Lactococcus lactis strain having anti-inflammatory activity, antioxidant activity, anticancer activity, antibacterial activity and anti-obesity activity and the use thereof' of the present invention. The problem to be solved

[0006] The present invention was derived from the above-mentioned needs, and the inventors obtained Lactococcus lactis (from healthy mice and humans) Lactococcus lactis The present invention was completed by isolating the SNU09LL strain (accession number: KCTC16429BP) and the Lactococcus lactis SNU10LL strain (accession number: KCTC16430BP) and confirming the excellent anti-inflammatory, antioxidant, anticancer, antibacterial, and anti-obesity activities of each isolated strain. means of solving the problem

[0007] To solve the above problem, the present invention relates to Lactococcus lactis having accession number KCTC16429BP, having anti-inflammatory activity, antioxidant activity, anticancer activity, antibacterial activity and anti-obesity activity ( Lactococcus lactis Provides any one strain selected from the group consisting of ) SNU09LL and Lactococcus lactis SNU10LL with accession number KCTC16430BP.

[0008] In addition, the present invention provides a health functional food composition for the prevention or improvement of cancer containing the strain, the culture medium thereof, the concentrate of the culture medium, or the dried product thereof as an active ingredient.

[0009] In addition, the present invention provides a pharmaceutical composition for the prevention or treatment of cancer containing the strain, the culture medium thereof, the concentrate of the culture medium, or the dried product thereof as an active ingredient.

[0010] In addition, the present invention provides a probiotic composition containing the strain, the culture medium thereof, the concentrate of the culture medium, or the dried product thereof as an active ingredient.

[0011] In addition, the present invention provides a postbiotic composition containing the strain, the culture medium thereof, the concentrate of the culture medium, or the dried product thereof as an active ingredient.

[0012] In addition, the present invention provides an anti-inflammatory composition containing the strain, the culture medium thereof, the concentrate of the culture medium, or the dried product thereof as an active ingredient.

[0013] In addition, the present invention provides an antioxidant food composition containing the strain, the culture solution thereof, the concentrate of the culture solution, or the dried product thereof as an active ingredient.

[0014] In addition, the present invention provides an antibacterial composition containing the strain, the culture solution thereof, the concentrate of the culture solution, or the dried product thereof as an active ingredient.

[0015] In addition, the present invention provides an anti-obesity composition containing the strain, the culture medium thereof, the concentrate of the culture medium, or the dried product thereof as an active ingredient. Effects of the invention

[0016] Lactococcus lactis of the present invention ( Lactococcus lactis Each of the SNU09LL and SNU10LL strains exhibits excellent anti-inflammatory, antioxidant, anticancer, antibacterial, and anti-obesity activities, and is a safe strain without hemolytic activity or bile salt degradation ability. Therefore, it can be utilized in compositions for the prevention, improvement, or treatment of cancer, probiotic compositions, anti-inflammatory compositions, antibacterial compositions, and anti-obesity compositions, and can be very useful in related industrial fields. Brief explanation of the drawing

[0017] Figure 1 shows the results of measuring the NO production inhibitory ability of each strain of Lactococcus lactis SNU09LL and SNU10LL of the present invention. Control: Group treated with medium without added strain. Figure 2 shows the results of measuring the DPPH radical scavenging activity of each strain of Lactococcus lactis SNU09LL and SNU10LL of the present invention. Ascorbic acid is the vitamin C-treated group used as a positive control. Figure 3 shows the analysis of the anticancer activity against colorectal cancer of each strain of Lactococcus lactis SNU09LL and SNU10LL of the present invention, where A is the result of measuring the cell viability of HT-29 cells and B is the result of measuring the cell viability of Caco-2 cells. Lactobacillus rhamnosus GG is the Lactobacillus rhamnosus GG strain treatment group used as a control group. FIG. 4 shows Staphylococcus aureus of each strain of Lactococcus lactis SNU09LL and SNU10LL of the present invention ( Staphylococcus aureus ), Listeria monocytogenes( Listeria monocytogenes ) and Salmonella enteritidis( Salmonella enteritidis This is the result of measuring the antibacterial activity against ). Figure 5 shows the analysis of the anti-obesity activity of each strain of Lactococcus lactis SNU09LL and SNU10LL of the present invention, where A is the result of calculating the adipocyte degradation ability by quantifying the Oil red O staining results, and B is a photograph observing the degree of adipocyte degradation through Oil red O staining. Control: Group treated with medium without added strain. Figure 6 shows the results of confirming the hemolytic activity and bile salt decomposition ability of each strain of Lactococcus lactis SNU09LL and SNU10LL of the present invention. Specific details for implementing the invention

[0018] To achieve the objective of the present invention, the present invention relates to Lactococcus lactis having accession number KCTC16429BP, having anti-inflammatory activity, antioxidant activity, anticancer activity, antibacterial activity, and anti-obesity activity ( Lactococcus lactis Provides any one strain selected from the group consisting of ) SNU09LL and Lactococcus lactis SNU10LL with accession number KCTC16430BP.

[0019] In the present invention, two strains possessing anti-inflammatory, antioxidant, anticancer, antibacterial, and anti-obesity activities were isolated from healthy mice and humans, and as a result of identification through 16S rRNA nucleotide sequence (SEQ No. 1, SEQ No. 2) analysis of each finally selected strain, it was found that both were Lactococcus lactis ( Lactococcus lactis It was confirmed that they were strains. The inventors named the two finally selected strains Lactococcus lactis SNU09LL and Lactococcus lactis SNU10LL, respectively, and deposited them with the Korea Culture Collection Center (KCTC) of the Korea Research Institute of Biotechnology and Bioengineering on August 8, 2025 (Deposit numbers: KCTC16429BP, KCTC16430BP).

[0020] In the strain according to the present invention, the anticancer effect may be anticancer against colorectal cancer, and the antibacterial activity may be Staphylococcus aureus ( Staphylococcus aureus ), Listeria monocytogenes( Listeria monocytogenes ) and Salmonella enteritidis( Salmonella enteritidis It may have antibacterial activity against ), but is not limited thereto.

[0021] Each of the Lactococcus lactis SNU09LL and SNU10LL strains of the present invention is characterized by having excellent nitric oxide production inhibition and free radical scavenging ability, having an ability to inhibit colon cancer cell proliferation, having excellent antibacterial activity against Staphylococcus aureus, Listeria monocytogenes and Salmonella enteritidis, having excellent adipocyte degradation ability, and having no hemolytic activity or bile salt degradation ability.

[0022] The present invention also provides a health functional food composition for the prevention or improvement of cancer, comprising the strain, the culture medium thereof, the concentrate of the culture medium, or the dried product thereof as an active ingredient.

[0023] In the health functional food composition for the prevention or improvement of cancer according to the present invention, the cancer may be colorectal cancer, but is not limited thereto.

[0024] The method of culturing each strain of Lactococcus lactis SNU09LL and SNU10LL of the present invention can be carried out according to methods commonly used in the art and is not limited to a specific method.

[0025] The "culture medium" of the present invention includes a processed product derived from the culture medium itself, such as the culture medium itself cultured in a liquid medium, or a filtrate (supernatant after centrifugation) from which strains have been removed by filtering or centrifuging the culture medium.

[0026] The health functional food composition of the present invention is preferably prepared in any one formulation selected from powder, granules, pills, tablets, capsules, candies, syrups, and beverages, but is not limited thereto. The health functional food composition of the present invention may be prepared by adding the active ingredient as is or by mixing it with other foods or food ingredients, and may be appropriately prepared according to conventional methods. Examples of foods to which the active ingredient may be added include any one form selected from caramel, meat, sausage, bread, chocolate, candies, snacks, confectionery, pizza, ramen, other noodles, chewing gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages, and vitamin complexes, and include all health functional foods in the conventional sense. That is, there are no special restrictions on the types of foods. The above health functional food composition may contain various nutritional supplements, vitamins, minerals (electrolytes), synthetic and natural flavoring agents, coloring agents and promoters (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc. In addition, it may contain fruit pulp for the production of natural fruit juices and vegetable beverages. The above ingredients may be used independently or in combination.

[0027] In addition, the health functional food composition of the present invention may contain various flavoring agents or natural carbohydrates as additional ingredients, and the natural carbohydrates are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. Although the proportion of the natural carbohydrates is not of great importance, it is preferable to include 0.01 to 0.04 g per 100 g of the composition of the present invention, and more preferably 0.02 to 0.03 g, but is not limited thereto. As sweeteners, natural sweeteners such as taumatin and stevia extract, or synthetic sweeteners such as saccharin and aspartame may be used.

[0028] The present invention also provides a pharmaceutical composition for the prevention or treatment of cancer containing the strain, the culture medium thereof, the concentrate of the culture medium, or the dried product thereof as an active ingredient.

[0029] In the pharmaceutical composition for the prevention or treatment of cancer according to the present invention, the cancer may be colorectal cancer, but is not limited thereto.

[0030] The pharmaceutical composition of the present invention may further comprise a pharmaceutically acceptable carrier, excipient, or diluent in addition to the active ingredient, and may be in various oral or parenteral formulations. When formulating, it is prepared using commonly used diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants. Solid formulations for oral administration include capsules, powders, granules, tablets, pills, etc., and these solid formulations are prepared by mixing at least one excipient, for example, starch, calcium carbonate, sucrose or lactose, gelatin, etc., with one or more compounds. In addition, lubricants such as magnesium stearate and talc are also used in addition to simple excipients. Liquid formulations for oral administration include suspensions, emulsions, syrups, and aerosols; in addition to commonly used simple diluents such as water and liquid laphine, they may contain various excipients, such as humectants, sweeteners, flavorings, and preservatives. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. Propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate may be used as non-aqueous solvents and suspension solvents. Witepsol, Macrogol, Tween 61, cocoa gelatin, laurin gelatin, and glycerogelatin may be used as bases for suppositories. For parenteral administration, it is preferable to select a method of administration via topical application or intraperitoneal, rectal, intravenous, intramuscular, subcutaneous, intrauterine dura mater, or intracerebrovascular injection.

[0031] The pharmaceutical composition according to the present invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" means an amount sufficient to treat a disease with a reasonable benefit / risk ratio applicable to medical treatment, and the level of the effective amount may be determined based on factors including the type and severity of the patient's disease, drug activity, sensitivity to the drug, time of administration, route of administration and elimination rate, duration of treatment, concurrently used drugs, and other factors well known in the medical field. The composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered as a single or multiple doses. It is important to administer an amount that obtains maximum effect with a minimum amount without side effects by taking all of the above-mentioned factors into account, and this can be easily determined by a person skilled in the art.

[0032] The present invention also provides a probiotic composition containing the strain, the culture medium thereof, the concentrate of the culture medium, or the dried product thereof as an active ingredient.

[0033] In the probiotic composition of the present invention, centrifugation or filtration may be performed to remove the culture medium from the strain culture solution and recover only the concentrated cells, and such steps may be performed by a person skilled in the art as necessary. The concentrated cells may be preserved by freezing or lyophilizing according to conventional methods so as not to lose their activity.

[0034] The term "probiotics" in this invention refers to live bacteria, that is, microorganisms that can remain and survive in the gastrointestinal tract when consumed by humans or animals, and refers to microbial preparations that have the effect of preventing or treating specific pathological conditions. In general, probiotics have the effect of treating and improving various symptoms caused by abnormal fermentation of the intestinal bacterial flora, and when administered to humans and animals, they densely settle and colonize the walls of the digestive tract in the intestines, thereby preventing harmful microorganisms from settling, and produce lactic acid to lower the intestinal pH, thereby inhibiting the proliferation of harmful microorganisms.

[0035] The probiotic composition of the present invention may be prepared and administered in various formulations and methods according to methods known in the art. For example, each strain of Lactococcus lactis SNU09LL and SNU10LL of the present invention, its culture medium, the concentrate of the culture medium, or the dried product thereof may be mixed with a carrier commonly used in the pharmaceutical field to be prepared and administered in the form of a powder, liquids and solutions, tablet, capsule, syrup, suspension, or granule. The carrier may be, for example, a binder, a lubricant, a disintegrant, an excipient, a solubilizer, a dispersant, a stabilizer, a suspending agent, a colorant, and a flavoring agent, but is not limited thereto. In addition, the dosage may be appropriately selected according to the absorption rate, inactivation rate, excretion rate of the active ingredient in the body, the age, sex, animal species, condition, and severity of the disease of the subject.

[0036] The present invention also provides a postbiotic composition containing the strain, the culture medium thereof, the concentrate of the culture medium, or the dried product thereof as an active ingredient.

[0037] Postbiotics are materials containing beneficial metabolites produced by probiotics during the fermentation process and components of microorganisms. Specifically, they include metabolites generated by probiotics, such as short-chain fatty acids, antimicrobial peptides, vitamins, extracellular polysaccharides, and cell wall components, as well as forms of live probiotics that have been treated to inactivate. Recent research results have reported that the efficacy of existing probiotics in improving various diseases is maintained in postbiotics at an equivalent level. Furthermore, postbiotics are attracting attention as a new alternative material capable of overcoming the limitations of safety, function, and stability associated with existing probiotic materials. Due to these advantages, postbiotics can be widely utilized as raw materials for health functional foods, cosmetics, and pharmaceuticals.

[0038] The present invention also provides an anti-inflammatory composition containing the strain, the culture medium thereof, the concentrate of the culture medium, or the dried product thereof as an active ingredient.

[0039] The anti-inflammatory composition of the present invention may be an anti-inflammatory pharmaceutical composition, an anti-inflammatory health functional food composition, or an anti-inflammatory cosmetic composition.

[0040] In addition, based on the anti-inflammatory effect described above, the composition of the present invention may be a composition for the prevention, treatment, or improvement of inflammatory diseases (e.g., a pharmaceutical composition for the prevention or treatment of inflammatory diseases, a health functional food composition for the prevention or improvement of inflammatory diseases, or a cosmetic composition for the prevention or improvement of inflammatory diseases (inflammatory skin diseases)).

[0041] In this context, inflammatory diseases refer to diseases characterized by inflammation as the primary lesion, and may be selected from the group consisting of, for example, gastritis, enteritis, hepatitis, pneumonia, nephritis, cystitis, arthritis, dermatitis, allergy, atopy, conjunctivitis, periodontitis, rhinitis, otitis media, and pharyngitis, but are not limited thereto.

[0042] The anti-inflammatory composition of the present invention may be administered orally or parenterally during clinical administration, and when administered parenterally, it may be administered by intraperitoneal injection, rectal injection, subcutaneous injection, intravenous injection, intramuscular injection, intrauterine dura injection, cerebral intravascular injection, or thoracic injection, and may be used in the form of a general pharmaceutical formulation.

[0043] The anti-inflammatory composition of the present invention may be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy, and biological response modulators.

[0044] The present invention also provides an antioxidant food composition containing the strain, the culture medium thereof, the concentrate of the culture medium, or the dried product thereof as an active ingredient.

[0045] The present invention also provides an antibacterial composition containing the strain, the culture solution thereof, the concentrate of the culture solution, or the dried product thereof as an active ingredient.

[0046] In the antibacterial composition of the present invention, the antibacterial agent is Staphylococcus aureus ( Staphylococcus aureus ), Listeria monocytogenes( Listeria monocytogenes ) or Salmonella enteritidis( Salmonella enteritidis It may be antibacterial against ), but is not limited to this.

[0047] The present invention also provides an anti-obesity composition containing the strain, the culture medium thereof, the concentrate of the culture medium, or the dried product thereof as an active ingredient.

[0048] The anti-obesity composition of the present invention may be an anti-obesity health functional food composition, an anti-obesity pharmaceutical composition, or an anti-obesity feed additive.

[0049] In addition, based on the anti-obesity effect described above, the composition of the present invention may be a composition for the prevention, treatment, or improvement of obesity (e.g., a health functional food composition for the prevention or improvement of obesity or a pharmaceutical composition for the prevention or treatment of obesity).

[0051] The present invention will be explained in detail below by way of examples. However, the following examples are merely illustrative of the present invention, and the scope of the present invention is not limited thereto.

[0053] Example 1. Isolation and Identification of Strains

[0054] Fecal samples from healthy mice and humans were inoculated into MRS (De Man Rogosa Sharpe) liquid medium and cultured at 37°C for 24 hours. Subsequently, the culture medium was plated onto MRS solid medium and cultured at 37°C for 24 hours; two single colonies formed were selected and purified. As a result of analyzing the nucleotide sequences of the 16S rRNA genes of the selected strains (Sequence No. 1, Sequence No. 2), Lactococcus lactis ( Lactococcus lactis It turned out to be, Lactococcus lactis SNU09LL, Lactococcus lactis They were named SNU10LL strains, respectively, and deposited at the Korea Culture Collection Center (KCTC) of the Korea Research Institute of Biotechnology and Bioengineering on August 8, 2025 (Deposit numbers: KCTC16429BP, KCTC16430BP).

[0055] After inoculating each strain of Lactococcus lactis SNU09LL and SNU10LL of the present invention into MRS liquid medium and anaerobic culturing at 37°C for 24 hours, 1×10 9The results were quantified to CFU / mL and further cultured at 37°C for 20 hours. Subsequently, the supernatant was removed by centrifugation at 4,000 rpm for 15 minutes, and the pellet was washed with PBS (Phosphate buffered saline). It was then suspended in antibiotic-free RPMI medium and anaerobically cultured at 37°C for 5 hours. Afterward, the supernatant (strain metabolites) obtained by centrifugation at 4,000 rpm for 15 minutes was filtered through a 0.22 µm filter and used for subsequent experiments.

[0057] Example 2. Analysis of anti-inflammatory activity

[0058] To analyze the anti-inflammatory activity of each strain of Lactococcus lactis SNU09LL and SNU10LL of the present invention, the inhibition rate of NO (Nitric oxide) production in RAW 264.7 cells with inflammation induced by LPS (lipopolysaccharide) was measured. RAW 264.7 cells were used in the experiment by culturing them in DMEM medium supplemented with 10% FBS and penicillin-streptomycin at 37°C and 5% CO2. RAW 264.7 cells were placed in a 24-well plate at a ratio of 1 x 10⁶ 5 After culturing cells / wells for 24 hours, each strain sample of the present invention (supernatant of Example 1) was pretreated with 10% LPS, then treated with LPS (1 μg / ml) and reacted for 24 hours. Subsequently, 100 μl of the cell culture medium was taken and reacted with grease reagent (1% sulfonic acid, 0.1% N-1-naphthyl ethylenediamine dihydrochloride, 2.5% phosphoric acid), and the absorbance was measured at 540 nm using a microplate reader. A group treated with MRS medium without added strains was used as a control.

[0059] As a result, it was confirmed that each strain of Lactococcus lactis SNU09LL and SNU10LL of the present invention has excellent activity in inhibiting NO production (Fig. 1).

[0061] Example 3. Antioxidant activity analysis

[0062] The antioxidant activity of each strain of Lactococcus lactis SNU09LL and SNU10LL of the present invention was analyzed according to the method of Blois et al. A 0.2 mM DPPH (2,2-diphenyl-1-picryl-hydrazyl) solution was prepared using methanol. The sample of each strain of the present invention (supernatant of Example 1) and the above DPPH reaction solution were mixed in equal volumes and reacted in a dark room for 30 minutes. Afterward, the absorbance was measured at 517 nm and calculated by substituting the values ​​into the equation below. Vitamin C was used as a positive control.

[0063] DPPH radical scavenging activity (%) = [1 - (A / B) × 100]

[0064] A: Absorbance of DPPH solution with sample added at 517 nm

[0065] B: Absorbance of DPPH solution without sample at 517 nm

[0066] As a result, it was confirmed that each strain of Lactococcus lactis SNU09LL and SNU10LL of the present invention has excellent DPPH radical scavenging ability (Fig. 2).

[0068] Examples 4. Analysis of Anticancer Activity

[0069] To analyze the anticancer activity of each strain of Lactococcus lactis SNU09LL and SNU10LL of the present invention, the viability of colorectal cancer cells [HT-29 (KCLB 30022), Caco-2 (KCLB 30038)] was measured. Colorectal cancer cells cultured in RPMI medium containing 10% FBS (Fetal Bovine Serum), 29.2 mg / mL glutamine, 10,000 units / mL penicillin, and 0.1 mg / mL streptomycin were placed in a 96-well plate at a concentration of 1 x 10⁶ 4Cells were dispensed at a density of cells / mL and cultured separately at 37°C and 5% CO2 for 24 hours. Subsequently, each strain sample of the present invention (supernatant of Example 1) was diluted to concentrations of 0–80% in antibiotic-free RPMI medium and treated separately for 24 hours. Then, the CCK-8 kit was dispensed at 10% of the total volume of each well and cultured for 2 hours. After measuring the absorbance at 450 nm, the cell viability (%) was calculated by substituting the values ​​into the following formula. Lactobacillus rhamnosus GG, known for its excellent anticancer activity ( Lactobacillus rhamnosus The GG strain was used as a control group.

[0070] Cell viability (%) = {(Absorbance of sample-treated group) / (Absorbance of control group)} × 100

[0071] As a result, it was confirmed that each strain of Lactococcus lactis SNU09LL and SNU10LL of the present invention had a superior inhibitory effect on colon cancer cell proliferation compared to the control group strain of Lactobacillus rhamnosus GG (Fig. 3).

[0073] Example 5. Antimicrobial Activity Analysis

[0074] 1.0×10 in TSB liquid medium 9 Pathogen quantified in CFU / mL [Staphylococcus aureus ( Staphylococcus aureus ), Listeria monocytogenes( Listeria monocytogenes ), Salmonella enteritidis( Salmonella enteritidis )] and 1.0×10 in MRS liquid medium 9 The Lactococcus lactis strain of the present invention [SNU09LL or SNU10LL], quantified in CFU / mL, was mixed in a volume ratio of 1:100 and co-cultured under anaerobic conditions at 37°C for 24 hours. Subsequently, each culture medium was serially diluted and cultured in selective agar media (mannitol salt medium for Staphylococcus aureus, PALCAM medium for Listeria monocytogenes, and XLT4 medium for Salmonella enteritidis) in which only each pathogen could grow, and the number of colonies was calculated. Each pathogen was cultured separately and used as a control.

[0075] As a result, it was confirmed that each strain of Lactococcus lactis SNU09LL and SNU10LL of the present invention has excellent antibacterial activity against Staphylococcus aureus, Listeria monocytogenes, and Salmonella enteritidis (Fig. 4).

[0077] Example 6. Analysis of anti-obesity activity

[0078] To analyze the anti-obesity activity of each strain of Lactococcus lactis SNU09LL and SNU10LL of the present invention, adipocyte degradation ability was measured. 3T3-L1 preadipocytes were placed in a 24-well plate in DMEM (Dulbecco Modified Eagle Medium) at a rate of 1 x 10⁶ 5 Cells were dispensed at a density of 1 / mL and cultured at 37°C and 5% CO2 for 2 days. Subsequently, differentiation was induced by treating with DMEM medium containing 0.5 μM IBMX (3-isobutyl-1-methylxanthine), 0.25 μM dexamethasone, and 10 μg / mL insulin. The DMEM medium was then replaced with fresh medium containing 10 μg / mL insulin every 2 days to allow the cells to mature into adipocytes for a total of 10 days. Afterward, the strain sample of the present invention (supernatant of Example 1) was applied and cultured for 24 hours, after which the degree of adipocyte degradation was measured by Oil red O staining. A group treated with medium without the strain was used as a control.

[0079] As a result, it was confirmed that each strain of Lactococcus lactis SNU09LL and SNU10LL of the present invention has excellent adipocyte degradation ability (Fig. 5).

[0081] Example 7. Confirmation of Safety

[0082] Each strain of Lactococcus lactis SNU09LL and SNU10LL of the present invention was cultured on blood agar medium (Blood TSA agar) and MRS solid medium containing 0.5% deoxycholic acid (TDCA) at 37°C for 24 to 48 hours, respectively, to observe whether a halo formed around the colony.

[0083] As a result, it was confirmed that each strain of Lactococcus lactis SNU09LL and SNU10LL of the present invention is a safe strain with γ-hemolytic activity that does not cause hemolysis and has no bile salt degradation ability (Fig. 6).

[0085] Based on the above results, it was found that each strain of Lactococcus lactis SNU09LL and SNU10LL of the present invention has excellent anticancer activity, anti-inflammatory activity, antioxidant activity, antibacterial activity, and anti-obesity activity, and is a safe strain with γ-hemolytic activity and no bile salt degradation ability.

[0086] Depository Name: Korea Research Institute of Biotechnology and Bioengineering Biological Resource Center (KCTC) Trustee Number: KCTC16429BP Date of Deposit: 2025-08-08 Depository Name: Korea Research Institute of Biotechnology and Bioengineering Biological Resource Center (KCTC) Trustee Number: KCTC16430BP Date of Deposit: 2025-08-08

Claims

Claim 1 Lactococcus lactis with accession number KCTC16429BP, having anti-inflammatory, antioxidant, anticancer, antibacterial, and anti-obesity activities ( Lactococcus lactis Any one strain selected from the group consisting of ) SNU09LL and Lactococcus lactis SNU10LL with accession number KCTC16430BP. Claim 2 In claim 1, the strain is characterized by having no hemolytic activity and bile salt decomposition ability. Claim 3 In claim 1, the antibacterial activity is Staphylococcus aureus ( Staphylococcus aureus ), Listeria monocytogenes( Listeria monocytogenes ) and Salmonella enteritidis( Salmonella enteritidis A strain characterized by having antibacterial activity against ). Claim 4 A health functional food composition for the prevention or improvement of colorectal cancer containing, as an active ingredient, a strain of any one of claims 1 to 3, a culture solution thereof, a concentrate of said culture solution, or a dried product thereof. Claim 5 A pharmaceutical composition for the prevention or treatment of colorectal cancer containing, as an active ingredient, a strain of any one of claims 1 to 3, a culture medium thereof, a concentrate of said culture medium, or a dried product thereof. Claim 6 delete Claim 7 A probiotic composition containing, as an active ingredient, a strain of any one of claims 1 to 3, a culture solution thereof, a concentrate of said culture solution, or a dried product thereof. Claim 8 A postbiotic composition containing, as an active ingredient, a strain of any one of claims 1 to 3, a culture solution thereof, a concentrate of said culture solution, or a dried product thereof. Claim 9 A health functional food composition for anti-inflammatory purposes containing, as an active ingredient, a strain of any one of claims 1 to 3, a culture solution thereof, a concentrate of said culture solution, or a dried product thereof. Claim 10 An antioxidant food composition containing, as an active ingredient, a strain of any one of claims 1 to 3, a culture solution thereof, a concentrate of said culture solution, or a dried product thereof. Claim 11 An antibacterial composition comprising as an active ingredient a strain of any one of claims 1 to 3, a culture solution thereof, a concentrate of said culture solution, or a dried product thereof, wherein the antibacterial agent is Staphylococcus aureus ( Staphylococcus aureus ), Listeria monocytogenes( Listeria monocytogenes ) or Salmonella enteritidis( Salmonella enteritidis An antimicrobial composition characterized by being antimicrobial against ). Claim 12 delete Claim 13 A health functional food composition for anti-obesity containing, as an active ingredient, a strain of any one of claims 1 to 3, a culture solution thereof, a concentrate of said culture solution, or a dried product thereof.

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