Faecalibacterium prausnitzii, postbiotic thereof, and use thereof

Abstract

Provided are a Faecalibacterium prausnitzii, a postbiotic thereof, and a use thereof. The Faecalibacterium prausnitzii is selected from a group consisting of an NCKUMM001 strain and an NCKUMT002 strain; the postbiotic of the Faecalibacterium prausnitzii is present in a conditioned medium thereof, and comprises an extracellular vesicle and a short-chain fatty acid. The Faecalibacterium prausnitzii and the postbiotic thereof have the effect of protecting the gut of an individual, slowing down inflammation, and regulating the expression of genes associated with lipid metabolism.

Description

Clostridium tenella, its postbiotics and uses thereof Technical Field

[0001] The present invention relates to Clostridium tenella, postbiotics thereof and uses thereof. Background Art

[0002] Probiotics generally refer to bacteria that are beneficial to organisms. It is currently believed that they are helpful for the intestinal health and even the immune system of organisms. Recent studies have pointed out that postbiotics, which are the products of probiotic metabolism, are important molecules for probiotics to achieve their beneficial effects. Postbiotics include extracellular vesicles, organic acids, short-chain fatty acids, exopolysaccharides, lipopolysaccharides, probiotic lysates, vitamins and amino acids, etc. Postbiotics can help the growth of probiotics or inhibit the growth of harmful microorganisms to improve the intestinal flora; postbiotics also have biological activities such as reducing inflammatory responses, anti-oxidation, and slowing down high blood sugar in organisms. In addition, more and more different bacteria have been proven to be helpful to the health of organisms, so the types of bacteria covered by probiotics are also becoming wider and wider.

[0003] However, probiotics of different strains, as well as probiotics of the same strain but different isolates, do not provide exactly the same benefits to organisms. Therefore, isolating various probiotics with different biological activities has always been an important research and development direction for researchers in related fields.

[0004] Summary of the Invention

[0005] The present invention relates to Faecalibacterium prausnitzii, postbiotics thereof, and uses thereof. The Faecalibacterium prausnitzii is selected from the group consisting of strains NCKUMT001 and NCKUMT002, wherein the NCKUMT001 strain has a deposit number of NITE BP-03861 and the NCKUMT002 strain has a deposit number of NITE BP-03862.

[0006] The NCKUMT001 in the present invention was deposited on March 20, 2023, at the National Institute of Technology, Japan, with the National Institute of Technology, Japan ...

[0007] The NCKUMT002 described in the present invention was deposited with the National Institute for Product Evaluation and Research (NPMD) on March 20, 2023, at the National Institute for Product Evaluation and Research (NIRT) ...

[0008] The present invention also relates to a composition containing postbiotics of Clostridium tenella NCKUMT001 or NCKUMT002.

[0009] Furthermore, the present invention also relates to the use of Clostridium truncatum and postbiotics derived from Clostridium truncatum in preparing compositions for anti-inflammation, intestinal protection, improved lipid metabolism, protection against intestinal infection-induced damage, alleviation of intestinal infection symptoms, and reduction of intestinal infection mortality. The compositions comprise at least one of Clostridium truncatum and postbiotics derived from Clostridium truncatum, and the compositions can be used to inhibit the expression of genes associated with inflammation, maintain intestinal cell integrity, and promote the expression of genes associated with lipid metabolism.

[0010] In one embodiment, the postbiotics of Clostridium tenella are isolated from the conditioned culture medium of the NCKUMT001 strain or the NCKUMT002 strain, including extracellular vesicles.

[0011] In one embodiment, the postbiotics of Clostridium tenella comprise short-chain fatty acids.

[0012] In one embodiment, the composition containing postbiotics of Clostridium tenella NCKUMT001 or NCKUMT002 further comprises at least one of a food or pharmaceutically acceptable carrier, excipient, adjuvant, antioxidant, or food additive.

[0013] In one embodiment, the inflammation-related genes include serum amyloid A1, interleukin-1β, macrophage inflammatory protein-2 (MIP-2), and interleukin-8.

[0014] In one embodiment, the lipid metabolism-related gene comprises the peroxisome proliferator-activated receptor γ gene.

[0015] In one embodiment, the intestinal bacteria metabolism genes include one or both of a secondary bile acid converting enzyme (HSDH) gene and a short-chain fatty acid converting enzyme gene.

[0016] In one embodiment, the Clostridium tenella postbiotics alleviate intestinal inflammation and damage to intestinal cell tight junctions caused by pathogen infection, protect against damage induced by intestinal infection, alleviate symptoms of intestinal infection, and reduce mortality from intestinal infection.

[0017] In one embodiment, the Clostridium tenella postbiotic promotes the expression of the tight junction protein (CLDN1) gene.

[0018] Thus, the Clostridium tenuis and postbiotics of Clostridium tenuis, such as its secretosomes, of the present invention have multiple functions, including alleviating inflammatory responses, protecting intestinal health, regulating lipid metabolism, protecting against damage induced by intestinal infection, alleviating symptoms of intestinal infection, and reducing mortality from intestinal infection. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1: Gram staining analysis photos and electron micrographs of exosomes of Clostridium tenella.

[0020] Figure 2: Phylogroup analysis of Clostridium tenella.

[0021] Figure 3: Expression analysis of short-chain fatty acid converting enzymes in Clostridium tenella.

[0022] Figure 4: Analysis of substances regulating inflammatory responses in Clostridium tenella and nutrient secretions of Clostridium tenella.

[0023] Figure 5: Analysis of the effects of nutrient secretions and exosomes of Clostridium tenella on cellular inflammatory substances, membrane potential, and tight junction protein (CLDN1) gene expression.

[0024] Figure 6: Analysis of the effects of postbiotics of Clostridium tenella on the survival rate and cecum length of experimental animals with intestinal infection.

[0025] Figure 7: Analysis of the effects of postbiotics from Clostridium tenella on inflammatory response substances in experimental animals with intestinal infection.

[0026] Figure 8: Analysis of the effects of postbiotics of Clostridium tenella on the number of intestinal flora, bile acid metabolism, and short-chain fatty acid metabolism gene expression.

[0027] Figure 9: Postbiotic effects of Clostridium tenella on experimental animals and analysis of gene expression related to lipid metabolism.

[0028] Figure 10: Analysis of the effects of postbiotics from Clostridium tenella on gastric parietal cell inflammation.

[0029] Biomaterial Deposit

[0030] (1) NCKUMT001 strain

[0031] Depository: NITE Patent Microorganisms Depositary National Institute of Technology and Evaluation (NPMD)

[0032] Address of the depository: Chiba, Japan (#122,2-5+8 Kazusakamatari, Kisarazu-shi, Chiba 292-0818, Japan)

[0033] Date of deposit: March 20, 2023

[0034] Accession number: NITE BP-03861

[0035] Classification name: Faecalibacterium prausnitzii

[0036] (2) NCKUMT002 strain

[0037] Depository: NITE Patent Microorganisms Depositary National Institute of Technology and Evaluation (NPMD)

[0038] Address of the depository: Chiba, Japan (#122,2-5+8 Kazusakamatari, Kisarazu-shi, Chiba 292-0818, Japan)

[0039] Date of deposit: March 20, 2023

[0040] Accession number: NITE BP-03862

[0041] Classification name: Faecalibacterium prausnitzii DETAILED DESCRIPTION

[0042] In order to provide a more complete and clear disclosure of the effects achievable by the technical means of the present invention, a detailed description is given below, and please refer to the disclosed drawings.

[0043] Among them, the postbiotics referred to in the present invention include substances with biological activity isolated from the conditioned culture medium of Clostridium tenella, including but not limited to cytosomes, proteins and / or short-chain fatty acids.

[0044] 1. Isolation of Clostridium tenella strains

[0045] The strains NCKUMT001 and NCKUMT002 tested in this example were both isolated from human fecal samples. They were cultured in BHIS (Brain-Heart Infusion Broth) at 37° C. in an anaerobic environment.

[0046] Furthermore, 16S rRNA sequencing of strains NCKUMT001 and NCKUMT002 revealed 98-99% similarity with the 16S rRNA of a standard strain of Clostridium truncatum. Therefore, strains NCKUMT001 and NCKUMT002 were determined to be Clostridium truncatum. The standard strains of Clostridium truncatum used in this experiment were Clostridium truncatum ATCC 27768 (hereinafter referred to as ATCC 27768) and Clostridium truncatum APC 918 / 95b (hereinafter referred to as APC 918 / 95b), respectively. The ATCC 27768 strain is designated ATCC 27768 by the American Type Culture Collection, and the APC 918 / 95b strain is designated JCM 39207 by the Japan Collection of Microorganisms. Please refer to (A) in Figure 1, which is a Gram-stained photograph of strain NCKUMT001, strain NCKUMT002, and strain ATCC27768. All three strains are Gram-positive bacteria with similar growth patterns.

[0047] Next, genomic deoxyribonucleic acid (DNA) was extracted from strains NCKUMT001 and NCKUMT002, and polymerase chain reaction (PCR) was performed using primers that specifically identify Clostridium tenella type I to determine the genotypes (Phylogroups) of the two strains. As shown in the agar gel electrophoresis photograph (A) in Figure 2 , strains ATCC27768 and NCKUMT002 belong to type I (Phylogroup I), while strain NCKUMT001 belongs to type II (Phylogroup II). Figure 2 (B) shows the results of random amplified polymorphic DNA polymerase chain reaction (PCR). This test uses primers targeting different bacterial genes to perform PCR, and the products are subjected to agarose gel electrophoresis to observe the DNA fragments amplified by each strain. As shown in Figure 2 (B), the DNA fragments amplified by strains NCKUMT002 and ATCC27768 are relatively similar, while the DNA fragment amplified by strain NCKUMT001 is different from those of the aforementioned two strains, confirming that strain NCKUMT001 and the other two strains are different types.

[0048] In addition, both the NCKUMT001 strain and the NCKUMT002 strain have been deposited at the NITE Patent Microorganisms Depositary National Institute of Technology and Evaluation, an independent administrative institution in Japan, and the deposit number of the NCKUMT001 strain is NITE BP-03861, and the deposit number of the NCKUMT002 strain is NITE BP-03862.

[0049] 2. Isolation and Analysis of Nutritional Secretions of Clostridium tenella

[0050] Seed cultures of strains NCKUMT001 and NCKUMT002 were first cultured in mBHIS (modified Brain-Heart Infusion Broth) medium under anaerobism for 48 hours. The culture broth was centrifuged at 4000 rpm for 10 minutes, and the supernatant was collected. This supernatant constitutes the conditioned medium of C. tenella and is subsequently referred to as "supplemental nutrient solution (SUP)." FIG. 1 (B) shows an electron microscopic photograph of extracellular vesicles (EVs) observed within the SUP of strain ATCC27768. Furthermore, as shown in Table 1, the concentrations of EVs and protein in the SUPs of strains ATCC27768, NCKUMT001, and NCKUMT002 were measured using a particle concentration assay and a qNano instrument. The results in Table 1 demonstrate that all three strains produce EVs and that the EVs contain protein.

[0051] Table 1

[0052] Please refer to Figure 3, which shows the analysis results of the relative expression levels of short-chain fatty acids (SCFAs) in the three strains detected by quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR). The results show that the SCFAs in strain NCKUMT001 are higher than those in strains ATCC27768 and NCKUMT002.

[0053] Please refer to Table 2, which shows the analysis results of the concentrations of medium and short-chain fatty acids in the nutrient secretions (SUP) of the NCKUMT001 and NCKUMT002 strains. The butyric acid content in the nutrient secretions (SUP) of the two strains is much higher than that of the blank culture medium; the formic acid and acetic acid contents in the nutrient secretions (SUP) of the NCKUMT001 strain are also much higher than those of the blank culture medium.

[0054] Table 2

[0055] Table 3 shows the minimum inhibitory concentration test results of different antibiotics for strains NCKUMT001 and NCKUMT002. The tested antibiotics include Ertapenem, Imipenem, Clindamycin, Metronidazole, and Ampicillin-Sulbactum. The results show that both strains have no resistance.

[0056] Table 3

[0057] 3. Biological activity test of NCKUMT001 strain and NCKUMT001 strain

[0058] (1) Inhibit inflammatory response

[0059] This experiment used a human colon cancer cell line (HT29). The cells were first stimulated with 100 mg / mL lipopolysaccharide (LPS) and 10 ng / mL interferon-γ (IFN-γ). One hour after stimulation, a nutrient secretion (SUP) containing 10% (v / v) NCKUMT001 was added to the cell culture medium. The cells were cultured for an additional hour, harvested, and RNA extracted. Real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) was then used to analyze the expression of interleukin-8 (IL-8) mRNA in the cells. As shown in Figure 4 (A), the IL-8 expression levels in the groups supplemented with the SUP were significantly lower than those in the group without the SUP. This result demonstrates that the SUP of the NCKUMT001 strain has the effect of reducing the expression of inflammatory substances.

[0060] Please also refer to Figure 4 (B), which shows that after the HT29 cell line was co-stimulated with LPS and IFN-γ, 5% to 40% (v / v) nutrient secretions (SUP) of the NCKUMT001 strain were added 1 hour after stimulation to observe the differences in cellular IL-8 mRNA expression levels. According to Figure 4 (B), the IL-8 expression level in the group with 5% (v / v) nutrient secretions was significantly lower than that in the group without added nutrient secretions. As the proportion of added nutrient secretions increased, the IL-8 expression level also decreased, showing a dose-dependent phenomenon.

[0061] Furthermore, in FIG4 (C), after HT29 cells were co-stimulated with LPS and IFN-γ, the cells were co-cultured with a bacterial solution containing 10% (v / v) NCKUMT001 cells, and the expression of IL-8 mRNA in cells of each group was observed. Among them, the low-concentration bacterial solution group used a bacterial solution with an absorbance value of "1" at a wavelength of 600nm (abbreviated as OD600), and the high-concentration bacterial solution group used a bacterial solution with an OD600 of "2". According to FIG4 (C), the IL-8 expression levels of both groups with added bacterial solution were lower than those of the group without added bacterial solution, indicating that the bacterial solution containing NCKUMT001 cells did not promote the inflammatory response, and even had the effect of slightly inhibiting the inflammatory response.

[0062] (2) Protection against Clostridium difficile-induced damage

[0063] Next, the protective effect of the nutrient secretion (SUP) or endosome (EV) of the NCKUMT001 strain on the ATCC43255 strain of Clostridium difficile (hereinafter abbreviated as CDI) was tested. In this experiment, HT29 cells were placed in a culture supernatant containing 10% (v / v) Clostridium difficile (CDI (SUP)) for 6 hours, and 20% (v / v) nutrient secretion (SUP) or endosome (EV) of the NCKUMT001 strain was added at the same time. The relative expression of IL-8 mRNA in the cells was observed. According to Figure 5 (A), the IL-8 expression level of the cell line added with the nutrient secretion (SUP) or endosome (EV) of the NCKUMT001 strain was significantly lower than that of the group without addition, that is, the nutrient secretion (SUP) or endosome (EV) of the NCKUMT001 strain does have the effect of reducing the expression of inflammatory substances.

[0064] 5 (B) shows a test of the effectiveness of Clostridium tenella in protecting cell membrane integrity. In the test, HT-29 cells were first cultured in the culture supernatant of Clostridium difficile (CDI(SUP)), and 20% (v / v) of the nutrient secretion (SUP) or endosomes (EV) of the NCKUMT001 strain were added at the same time and co-cultured for 2 hours. Then, the cells were stained with FITC (Fluorescein isothiocyanate) fluorescent dye. If the cell membrane is damaged, the fluorescent dye will enter the cell and stain the cell. The stained cells were collected and the fluorescence intensity of the cells in each group was measured. The stronger the fluorescence intensity detected in the group of cells, the more serious the cell membrane damage in the group of cells. According to (B) in Figure 5, the fluorescence intensity of the cells in the groups treated with the nutrient secretions (SUP) or endosomes (EV) of the NCKUMT001 strain was significantly lower than that of the untreated group, and the fluorescence intensity of the endosome (EV) treated group was the lowest among the three groups, indicating that the nutrient secretions (SUP) or endosomes (EV) of the NCKUMT001 strain can protect the cell membrane from cell membrane damage caused by Clostridium difficile.

[0065] In addition, Figure 5 (C) shows the expression of claudin (CLDN1) mRNA, a protein related to tight junctions between cells, in each group under the same treatment. Please refer to Figure 5 (C). The expression of claudin in cells treated with nutrient secretions (SUP) or endosomes (EV) of the NCKUMT001 strain was significantly higher than that in the untreated group, indicating that the nutrient secretions (SUP) or endosomes (EV) of Clostridium teneis have the effect of increasing the expression of claudin and thereby promoting tight junctions between cells.

[0066] (3) Animal experimental models

[0067] In this study, C57BL / 6J mice were used as an animal model to observe the effects of Clostridium tenella on intestinal health. Please refer to Figure 6 (A) for details. The start date of this experiment is defined as "Day 0," the day before the experiment is defined as "Day -1," the day after the experiment is defined as "Day 1," and so on. The mice were divided into the following groups:

[0068] (1) Blank control group: experimental mice did not receive any treatment at all;

[0069] (2) CDI group: Five days before the start of the experiment, the experimental mice were given antibiotics orally (antibiotic types and doses used: Kanamycin 0.4 mg / mL; Gentamycin 0.0035 mg / mL; Colistin 0.057 mg / mL; Metronidazole 0.215 mg / mL; Vancomycin 0.045 mg / mL) to destroy the normal intestinal flora of the experimental mice. After the start of the experiment, the mice were given 10 4 CFU / ml of Clostridium difficile spores were used to infect mice;

[0070] (3) CDI+EV group: 5 days before the start of the experiment, the experimental mice were given 200uL of NCKUMT001 strain EVs at a concentration of 10 11 / ml, and after infection with Clostridium difficile, 200uL of NCKUMT001 strain secretosomes were still administered daily until the end of the experiment.

[0071] On day 3 after infection, the surviving mice were sacrificed and their intestinal tissues were collected for subsequent analysis.

[0072] Please refer to Figure 6 (B) for the analysis of the survival rates of experimental mice in each group. No experimental mice in the blank control group had died at the end of the experiment, so their survival rate was 100%. On the third day after infection, the survival rate of experimental mice in the CDI group was 60%, while the survival rate of experimental mice in the CDI+EV group was the same as the blank control group, at 100%. This result shows that the exosomes of the NCKUMT001 strain can indeed prevent the death of experimental mice caused by infection with Clostridium difficile.

[0073] Figure 6 (C) shows the analysis results of the intestinal length of each group of experimental mice. In this experiment, the small intestine length of the experimental mice was measured. Compared with the blank control group, the large intestine length of the experimental mice in the CDI group was significantly decreased, while the intestinal length of the experimental mice in the CDI+EV group was significantly increased compared with the CDI group. This result shows that the exosomes of the NCKUMT001 strain can effectively slow down the intestinal damage caused by bacterial infection in mice.

[0074] Please refer to Figure 7 (A) and Figure 7 (B), which respectively show the expression levels of serum amyloid A1 (SAA1) and interleukin-1β (IL-1β) in the intestinal tissues of experimental mice. According to the experimental results, the expression levels of SAA1 and IL-1β in the intestinal tissues of experimental mice in the CDI group were significantly higher than those in the blank control group. However, the expression levels of these two cytokines in the intestinal tissues of experimental mice in the CDI+EV group were significantly lower than those in the CDI group. This result shows that the secretosomes of Clostridium tenella effectively reduce the expression of inflammatory proteins in intestinal cells.

[0075] (4) Clostridium tenella affects intestinal bacterial metabolism

[0076] The purpose of the following experiment was to test the biological activity of Clostridium tenella in relation to intestinal microbial metabolism. This experiment utilized an in vitro fermentation system developed by the present inventors. Human intestinal bacteria were cultured within this system to create a bacterial flora composition that simulated the human intestinal microbiome (hereinafter referred to as the intestinal flora composition). The expression of intestinal microbial-related metabolic genes was then observed under different conditions.

[0077] In this experiment, three groups were divided into: (1) blank control group, in which the intestinal flora composition in the in vitro fermentation system did not receive any treatment; (2) antibiotic group, in which 150 mg / mL clindamycin was added to the culture medium of the intestinal flora composition in the in vitro fermentation system and cultured together for 1 day; and (3) SUP group, in which 150 mg / mL clindamycin antibiotic and 10% (v / v) nutrient secretion (SUP) of the NCKUMT001 strain were added to the culture medium of the in vitro fermentation system for 1 day. After the experiment, DNA of the intestinal flora composition in each group was collected and the number of bile acid and short-chain fatty acid metabolism-related genes in the samples was detected and analyzed by real-time quantitative polymerase chain reaction.

[0078] Please refer to (A) in Figure 8, which is an analysis of the number of genes of intestinal bacteria in each group. Compared with the blank control group, the number of genes of intestinal bacteria in the group treated with antibiotics decreased significantly, but the number of genes of intestinal bacteria in the SUP group was significantly higher than that in the antibiotic group. This result shows that the nutrient secretion (SUP) of the NCKUMT001 strain can slow down the reduction of intestinal bacteria caused by antibiotics.

[0079] Please see Figure 8 (B), which shows the gene count analysis results for the secondary bile acid converting enzyme (hydroxysteroid dehydrogenase) in the intestinal flora composition of each group. Compared with the blank control group, the gene count of the secondary bile acid converting enzyme in the intestinal flora composition of the antibiotic-treated group decreased significantly, but the gene count of the secondary bile acid converting enzyme in the intestinal flora composition of the SUP group was higher than that of the antibiotic group.

[0080] Please refer to (C) in Figure 8, which is the gene quantity analysis results of the short-chain fatty acid convertase in the intestinal flora composition of each group; compared with the blank control group, the gene quantity of the short-chain fatty acid convertase in the intestinal flora composition of the antibiotic-treated group was significantly decreased, but the expression level of the intestinal flora composition of the SUP group was significantly higher than that of the antibiotic group.

[0081] The results in FIG8 demonstrate that the Clostridium tenella of the present invention can reduce the death of intestinal flora caused by antibiotic use, thereby restoring the metabolic function of intestinal bacteria damaged by antibiotic use, thereby promoting the ability of intestinal flora to metabolize bile acid and short-chain fatty acids, thereby maintaining intestinal health and regulating intestinal inflammatory responses.

[0082] Please refer to Figure 9 again, which shows the effect of Clostridium tenella nutrient secretions (SUP) on protecting cells from Clostridium difficile infection using HT-29 cells. The experiment included four groups: (1) a blank control group, in which the cells did not receive any treatment; (2) a CDI group, in which the cells were infected with Clostridium difficile at a multiplicity of infection (MOI) of 5; (3) a SUP group, in which the cells were infected with Clostridium difficile and cultured in a culture medium containing 10% (v / v) nutrient secretions (SUP) of the NCKUMT001 strain for 4 h; and (4) an EV group, in which the cells were infected with Clostridium difficile and cultured in a culture medium containing 10% (v / v) endosomes (EV) of the NCKUMT001 strain for 4 h.

[0083] The results in Figure 9 show that in cells infected with Clostridium difficile, the expression of peroxisome proliferator-activated receptor-γ (PPAR-γ) decreased significantly, reaching levels significantly lower than those in the blank control group. However, PPAR-γ levels in the SUP and EV groups significantly rebounded, even reaching levels higher than those in the blank control group. PPAR-γ is a key factor in regulating the human immune response. In addition to its ability to mitigate inflammation, it also promotes fat metabolism and reduces body weight.

[0084] (5) Clostridium tenella regulates inflammatory response

[0085] This experiment was conducted using GES-1 gastric epithelial cells. The GES-1 gastric epithelial cell line was first stimulated with lipopolysaccharide, and then the cells were co-cultured with NCKUMT001 bacterial solution (hereinafter referred to as the "bacterial solution group") or NCKUMT001 exosomes (hereinafter referred to as the "EV group"). The IL-8 expression level of cells in each group was observed one day after culture. The amount of NCKUMT001 added to the bacterial solution group was an MOI (multiplicity of infection) of 100, while the concentration of exosomes added to the EV group was 10 4 granular secretosomes / cell; please refer to Figure 10, Figure 10 (A) is the analysis result of IL-8 concentration in the cell culture supernatant, and Figure 10 (B) is the analysis result of the relative expression of IL-8 mRNA in cells of each group. According to Figure 10 (A) and (B), lipopolysaccharide stimulates cells to express IL-8, but the IL-8 mRNA expression and secretion of cells in the bacterial solution group and EV group are significantly reduced, that is, when the cells are co-cultured with NCKUMT001 bacterial solution or co-cultured with NCKUMT001 secretosomes, the lipopolysaccharide-induced IL-8 mRNA expression and IL-8 protein expression are inhibited, which also means that NCKUMT001 bacterial solution and its secretosomes not only do not promote inflammatory response, but may even significantly inhibit inflammatory response.

[0086] In summary, the present invention discloses Clostridium tenuis, its postbiotics, and uses thereof, comprising two novel Clostridium tenuis isolates. The nutrient cultures and secretions of these two strains have the efficacy of reducing gastrointestinal inflammation, maintaining gastrointestinal health, and regulating lipid metabolism. In embodiments of the present invention, the nutrient secretions of the Clostridium tenuis strains contain bioactive postbiotics, such as secretions or various short-chain fatty acids, which protect the integrity of intestinal cell membranes and promote the expression of genes related to lipid metabolism, thereby regulating lipid metabolism in an organism.

[0087] In summary, the Clostridium tenella, its postbiotics, and their uses of the present invention can indeed achieve the intended efficacy through the disclosed embodiments. Furthermore, the present invention, which was not disclosed prior to the application, fully complies with the provisions and requirements of the Patent Law. However, the above description is merely a preferred embodiment of the present invention and is not intended to limit the scope of protection of the present invention. Other equivalent variations or modifications made by those skilled in the art within the scope of the present invention should be considered to fall within the scope of the present invention.

Claims

1. A Faecalibacterium prausnitzii selected from the group consisting of Faecalibacterium prausnitzii strain NCKUMT001 and Faecalibacterium prausnitzii strain NCKUMT002, wherein the NCKUMT001 strain has a deposit number of NITE BP-03861 and the NCKUMT002 strain has a deposit number of NITE BP-03862.

2. A postbiotic of Faecalibacterium prausnitzii, isolated from the conditioned culture medium of Faecalibacterium prausnitzii strain NCKUMT001 or Faecalibacterium prausnitzii strain NCKUMT002, wherein the NCKUMT001 strain has a deposit number of NITE BP-03861 and the NCKUMT002 strain has a deposit number of NITE BP-03862.

3. The postbiotic of Clostridium tumefaciens according to claim 2, comprising a protein secreted by Clostridium tumefaciens. The postbiotic of Clostridium tenella according to claim 2 , comprising extracellular vesicles. The postbiotic of Clostridium tenella according to claim 2 , comprising short-chain fatty acids.

6. A composition comprising Clostridium truncatum or at least one of its postbiotics, wherein the Clostridium truncatum is selected from the group consisting of Clostridium truncatum NCKUMT001 strain or Clostridium truncatum NCKUMT002 strain, wherein the NCKUMT001 strain has a deposit number of NITE BP-03861, and the NCKUMT002 strain has a deposit number of NITE BP-03862.

7. The composition of claim 6, wherein the postbiotics of Clostridium tumefaciens comprise conditioned medium of Clostridium tumefaciens.

8. The composition of claim 6, wherein the postbiotics of Clostridium tenella comprise exosomes.

9. The composition of claim 6, wherein the postbiotics of Clostridium tenella comprise short-chain fatty acids.

10. The composition of claim 6, comprising at least one of a food or a pharmaceutically acceptable carrier, excipient, adjuvant, antioxidant or food additive.

11. A use of at least one of Clostridium truncatum or a postbiotic of Clostridium truncatum for preparing a composition for protecting intestinal health, wherein the composition inhibits the expression of genes associated with gastrointestinal inflammation, maintains intestinal cell integrity, promotes the expression of genes associated with lipid metabolism, promotes the expression of genes associated with fatty acid metabolism, promotes the expression of genes associated with bile acid metabolism, and reduces antibiotic-induced intestinal flora death, wherein the Clostridium truncatum is selected from the group consisting of Clostridium truncatum NCKUMT001 and Clostridium truncatum NCKUMT002, wherein the NCKUMT001 strain has a deposit number of NITE BP-03861, and the NCKUMT002 strain has a deposit number of NITE BP-03862.

12. The use according to claim 11, wherein the inflammation-related gene comprises serum amyloid A1, interleukin-1β, and interleukin-8, and wherein the lipid metabolism-related gene comprises the peroxisome proliferator-activated receptor γ gene, the fatty acid metabolism-related gene comprises the short-chain fatty acid convertase gene, and the bile acid metabolism-related gene comprises the secondary bile acid convertase gene.

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