Intestinal microbiome biomarker composition for distinguishing equol producers and use thereof

Abstract

The present invention relates to: an intestinal microbiome biomarker composition for distinguishing equol producers; and use thereof. Intestinal microbiome biomarkers are differently expressed between equol producers and non-producers, and thus equol producers can be distinguished by confirming the expression of the biomarkers. Therefore, it is expected that the intestinal microbiome biomarkers for distinguishing equol producers, of the present invention, can be used to simply distinguish between equol producers and non-producers.

Description

Intestinal microbial biomarker composition for determining equal producers and use thereof

[0001] The present invention relates to a gut microbiome biomarker composition for identifying equal-producer genes and its use. This application claims priority to Korean Patent Application No. 10-2024-0089727, filed July 8, 2024, the entire disclosure of which is incorporated herein by reference.

[0002] The probiotics market is booming, fueled by research showing a close connection between gut microbiota and immunity and disease. Gut microbiota are microorganisms residing within the human digestive tract, playing a crucial role in diverse physiological functions, including digestion, the immune system, and metabolism. The composition of the gut microbiota is determined by lifestyle, diet, and genetics, resulting in individual differences in gut microbiota composition.

[0003] The accelerated aging of the population and the spread of the self-medication trend are driving the increase in domestic and international consumption of health functional foods. While people purchase and consume health functional foods for health promotion, fatigue relief, and disease prevention, individual differences in metabolism lead to differences in actual benefits. The composition of the gut microbiome plays a key role in these differences. Gut microbiomes contain enzymes that ferment indigestible foods, thereby influencing the health of the host. Consequently, the products obtained from food intake vary depending on the composition of an individual's gut microbiome. Representative examples include urolithin, equal, and short-chain fatty acids.

[0004] Soybeans have long been used as a raw material for various traditional fermented foods, and are known to exhibit beneficial activities for humans when broken down through the fermentation process by various microorganisms. Daidzin, genistin, and glycitin are known to be the main isoflavones found in soybeans. In addition, daidzein, genistein, glycitein, and equol are produced during the soybean fermentation process and are known to be digested and absorbed more quickly in the body and enhance physiological activity. Among them, equol is a major metabolite of isoflavones, a major soybean substance, and is a functional substance produced through the metabolism of intestinal microorganisms.

[0005] While accumulated research suggests that EQUAL has a positive effect on various diseases, including cardiovascular disease and arteriosclerosis, it is reported that only 30-40% of the population can produce it. This suggests that gut microbes are crucial for EQUAL production, and the effects of EQUAL are expected to vary from person to person, depending on the presence and abundance of EQUAL-producing gut microbes.

[0006] Equal producers are entities capable of producing Equals through gut microbes. Equal production can be closely linked to an individual's health, so identifying them is a crucial step in developing personalized health management and treatments. Identifying Equal producers requires accurately analyzing the presence and activity of specific gut microbes associated with Equal production.

[0007] Previously, to identify the equator, each subject had to personally consume a health functional food containing soy isoflavones and analyze the precursors and metabolites in their feces before and after consumption. However, this clinical trial process was significantly costly and time-consuming.

[0008] Accordingly, the present invention proposes a biomarker composition using intestinal microorganisms and a method for providing information for identifying equal producers using the same.

[0009] The present inventors analyzed the differences in the gut microbiomes of equal producers and non-equal producers and confirmed that there were differences in gut microbial diversity. Accordingly, they confirmed that gut microbiota selected using the Boruta method can be used as biomarkers for identifying equal producers, and based on this, completed the present invention.

[0010] Accordingly, the purpose of the present invention is to provide an intestinal microbial biomarker composition for determining an equal producer, a composition for determining an equal producer, a kit for determining an equal producer, and a method for providing information for determining an equal producer, thereby enabling easy determination of whether an equal producer is present without a clinical trial process.

[0011] However, the technical problems to be solved by the present invention are not limited to the problems mentioned above, and other problems not mentioned can be clearly understood by a person having ordinary skill in the technical field to which the present invention belongs from the description below.

[0012] The terminology used herein is for the purpose of description only and should not be construed as limiting. The singular expression includes the plural expression unless the context clearly indicates otherwise. In this specification, the terms "comprises" or "has" and the like are intended to specify the presence of a feature, number, step, operation, component, part, or combination thereof described in the specification, but should be understood to not preclude the presence or addition of one or more other features, numbers, steps, operations, components, parts, or combinations thereof.

[0013] Unless otherwise defined, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by a person of ordinary skill in the art to which the embodiments pertain. Terms defined in commonly used dictionaries should be interpreted as having a meaning consistent with their meaning in the context of the relevant technology, and shall not be interpreted in an idealized or overly formal sense unless explicitly defined herein.

[0014] Hereinafter, the present invention will be described in more detail.

[0015] The present invention relates to a method for treating Peptostreptococcaceae, Christensenellaceae, Streptococcaceae, Leuconostocaceae, Eggerthella, Adlercreutzia, Family XIII UCG001, Intestinibacter, Ruminococcus, NK4A214 group, UCG010, Christensenellaceae R7 group, Streptococcus, Eubacterium coprostanoligenes group, Eubacterium ruminantium group, Catenibacillus, Provided is an intestinal microbial biomarker composition for determining equal producers, comprising as an active ingredient one or more intestinal microorganisms selected from the group consisting of Leuconostoc, Roseburia, and Blautia.

[0016] In the present invention, the "equal" is a substance produced by the conversion of soybean isoflavone by intestinal microorganisms, and has various health benefits, including cardiovascular and neurodegenerative diseases, but it is known that only about 30-40% of people produce equal. The equal has a molecular weight of 242.27 g / mol and a molecular formula of C 15 H 14 O3 has a structure represented by the following chemical formula 1.

[0017] [Chemical Formula 1]

[0018]

[0019] In the present invention, the “soy isoflavone” is a phytochemical naturally present in soybeans and is a type of phytoestrogen. The isoflavones contained in soybeans are structurally similar to the female hormone estrogen and have similar biological actions, so they are also called phytoestrogens. Chemically, soy isoflavone refers to a general term for flavonoid compounds that are abundantly contained in the hypocotyl region (bean germ) of soybeans. In the present invention, the “composition” is a biomarker composition for determining whether or not an equal producer is present, and may specifically include one or more intestinal microorganisms selected from the group consisting of the intestinal microorganisms as an active ingredient, but is not limited thereto.

[0020] In the present invention, the above “biomarker” refers to molecular information derived from DNA, RNA, metabolites, proteins, and protein fragments, etc., and is an indicator that can detect changes in the body caused by the occurrence of a disease, etc., and is widely used in the development of new drugs related to the occurrence and progression of diseases, in vitro molecular diagnostic technology that detects diseases early in vitro and observes their prognosis, personalized medical technology that determines the individual characteristics of biomarkers that react to specific drugs, and ubiquitous healthcare systems that create a convenient treatment environment for patients. Since a biomarker appears differently in a living organism depending on the type, occurrence, and progression of the disease, it is a substance that serves as an indicator in blood or body fluid that allows for objective measurement of the presence of a specific disease or drug response status, and it plays a role in enabling the early diagnosis of a disease simply by analyzing this blood or body fluid.

[0021] In the present invention, the "gut microbial biomarker" refers to an indicator that can detect changes in the body using intestinal microorganisms. The intestinal microbial biomarker is intended to identify equal producers, and may refer to, but is not limited to, intestinal microorganisms or communities that exhibit a specifically higher or lower level of expression compared to equal non-producers.

[0022] In the present invention, the “intestinal microorganisms” may be selected by calculating feature importance based on a random forest using the Boruta technique, but are not limited thereto.

[0023] In the present invention, the "Boruta technique" is a wrapper algorithm for variable selection, used to select important features. The Boruta technique evaluates the importance of all variables and clearly distinguishes between important and unimportant variables.

[0024] In the present invention, the intestinal microbiota biomarker selected from the equal generator may be evaluated to calculate the AUC (Area Under the Curve) value for the ROC (Receiver Operation Characteristic) curve, but is not limited thereto.

[0025] In the present invention, at least one intestinal microorganism selected from the group consisting of Christensenellaceae, Leuconostocaceae, Peptostreptococcaceae, Adlercreutzia, Catenibacillus, Christensenellaceae R7 group, Eubacterium coprostanoligenes group, Eubacterium ruminantium group, Family XIII UCG001, Intestinibacter, Leuconostoc, NK4A214 group, Roseburia, Ruminococcus, and UCG010 is equal It may be upwardly expressed in equal generators compared to non-generators, but is not limited thereto.

[0026] In the present invention, one or more intestinal microorganisms selected from the group consisting of Streptococcaceae, Blautia, Eggerthella, and Streptococcus may be down-expressed in equal producers compared to equal non-producers, but is not limited thereto.

[0027] In addition, the present invention relates to a method for treating Peptostreptococcaceae, Christensenellaceae, Streptococcaceae, Leuconostocaceae, Eggerthella, Adlercreutzia, Family XIII UCG001, Intestinibacter, Ruminococcus, NK4A214 group, UCG010, Christensenellaceae R7 group, Streptococcus, Eubacterium coprostanoligenes group, Eubacterium ruminantium group, A composition for determining an equal producer is provided, characterized in that it includes a preparation capable of detecting one or more intestinal microorganisms selected from the group consisting of Catenibacillus, Leuconostoc, Roseburia, and Blautia.

[0028] In the present invention, the “detectable agent” refers to a substance that can be used to detect the presence of the intestinal microorganism, which is an equal-producer discrimination marker, in a sample. The “detectable agent” may be, but is not limited to, a primer, a probe, an aptamer, an avidity multimer, a peptidomimetic, or an antibody.

[0029] In the present invention, the “detection” may be performed through a metagenome such as next generation sequencing analysis, but is not limited thereto.

[0030] In the present invention, the "primer" refers to a single-stranded oligonucleotide sequence complementary to a nucleic acid strand to be amplified, and can act as a starting point for the synthesis of a primer extension product. The appropriate length of the primer may vary depending on the temperature and the intended use of the primer. In addition, the sequence of the primer does not need to be completely complementary to a portion of the template sequence, and it is sufficient if it has sufficient complementarity within a range where it can hybridize with the template and perform its own function.

[0031] In the present invention, the primer sequence may be modified using a label capable of directly or indirectly providing a detectable signal. The primer may include a label detectable using spectroscopic, photochemical, biochemical, immunochemical, or chemical means. Useful labels include 32P, fluorescent dyes, electron-dense reagents, enzymes (typically used in ELISA), biotin or haptens, and proteins to which antiserum or monoclonal antibodies are available.

[0032] In the present invention, the “probe” means a nucleic acid fragment corresponding to several to several hundred bases that can specifically bind to DNA or RNA, and can be produced in the form of an oligonucleotide probe, a single-stranded DNA probe, a double-stranded DNA probe, or an RNA probe.

[0033] In the present invention, the "aptamer" refers to a DNA, RNA, or derivative thereof, measuring approximately 20 to 100 nucleotides in size, that possesses a stable tertiary structure and high affinity and specificity for a specific target molecule. Aptamers, like antibodies, possess the property of binding strongly and specifically to target proteins, and are therefore used in place of antibodies for diagnostic or therapeutic purposes.

[0034] In the present invention, the "avimer" refers to two or more peptides of 30 to 35 amino acids in length linked together using a linker. Avimer is an abbreviation for avidity multimer, which refers to increasing affinity by increasing the number of peptides that bind to a target antigen. Avidity refers to the effect of the number of binding sites on binding strength in protein-protein binding. The affinity for the target can increase depending on the number of binding domains.

[0035] In the present invention, the “peptide mimic” refers to a chemical structure that can replace a natural peptide in interacting with a receptor or enzyme.

[0036] In the present invention, the “antibody” refers to something that can detect a microorganism using an immunological method based on an antigen-antibody reaction. Analysis methods for this purpose include, but are not limited to, Western blot, ELISA (enzyme linked immunosorbent assay), RIA (Radioimmunoassay), radioimmunodiffusion, Ouchterlony immunodiffusion, rocket immunoelectrophoresis, tissue immunostaining, immunoprecipitation assay, complement fixation assay, FACS (Fluorescence activated cell sorter), protein chip, etc.

[0037] In addition, the present invention provides a kit for determining an equal generator, comprising the composition for determining an equal generator.

[0038] In the present invention, the “kit” may be, but is not limited to, an RT-PCR (Reverse transcription polymerase chain reaction) kit, a DNA chip kit, an ELISA (Enzymelinked immunosorbent assay) kit, a protein chip kit, a rapid kit, or an MRM (Multiple reaction monitoring) kit.

[0039] In the present invention, the kit may include not only a primer, probe, aptamer, avimer, peptide mimic, and antibody detection agent for detecting the intestinal microorganism, but also one or more additional component compositions, solutions, or devices suitable for the analysis method.

[0040] In the present invention, the kit including the primer for detecting the intestinal microorganisms may be a kit including essential elements for performing an amplification reaction such as PCR. For example, the PCR kit may include, but is not limited to, a test tube or other appropriate container, a reaction buffer, deoxynucleotides (dNTPs), an enzyme such as Taq polymerase reverse transcriptase, DNase, RNAse inhibitor, DEPC water, sterile water, etc.

[0041] In the present invention, the kit can be applied to a sample collected from the intestines, and further, the kit can further include instructions for collecting a sample from the intestines of a subject to determine the equal generator, but is not limited thereto.

[0042] In addition, the present invention provides a method for isolating Peptostreptococcaceae, Christensenellaceae, Streptococcaceae, Leuconostocaceae, Eggerthella, Adlercreutzia, Family XIII UCG001, Intestinibacter, Ruminococcus, NK4A214 group, UCG010, Christensenellaceae R7 group, Streptococcus, Eubacterium coprostanoligenes group, Eubacterium ruminantium group, A method for providing information for determining an equal producer is provided, comprising: a step of detecting one or more intestinal microorganisms selected from the group consisting of Catenibacillus, Leuconostoc, Roseburia, and Blautia; and a step of comparing the expression level of the intestinal microorganisms with a control group.

[0043] In the present invention, the “individual” means any living organism that is divided into an equalizer or a non-equalizer, or whether soybean isoflavones are converted into equalizers in the intestines, and specific examples thereof include mammals, birds, farmed fish, etc., including mice, monkeys, cows, pigs, mini-pigs, livestock, and humans, without limitation.

[0044] In the present invention, the "sample" means a material derived from the individual, and may be specifically cells, urine, feces, etc., but may be Peptostreptococcaceae, Christensenellaceae, Streptococcaceae, Leuconostocaceae, Eggerthella, Adlercreutzia, Family XIII UCG001, Intestinibacter, Ruminococcus, NK4A214 group, UCG010, Christensenellaceae R7 group, Streptococcus, Eubacterium coprostanoligenes group, Eubacterium There is no limitation on the type of intestinal microorganisms that can be detected, as long as they are Eubacterium ruminantium group, Catenibacillus, Leuconostoc, Roseburia, or Blautia.

[0045] In the present invention, the “control group” may be a sample of an individual determined to be an equal producer or a non-producer, and the control group is for comparing the expression level of intestinal microorganisms detected from a sample of an individual not determined to be an equal producer or a non-producer, and refers to an individual that can be determined as an equal producer or a non-producer based on the comparison result. Specifically, if the expression level of intestinal microorganisms in a sample of an undefined individual is significantly higher or lower than the expression level of intestinal microorganisms in an equal non-producer, the undefined individual can be determined as an equal producer. In addition, if the expression level of intestinal microorganisms in a sample of an undefined individual is significantly higher or lower than the expression level of intestinal microorganisms in an equal non-producer; and if the expression level of intestinal microorganisms in a sample of an undefined individual is similar to the expression level of intestinal microorganisms in an equal producer; the undefined individual can be determined as an equal producer, but is not limited thereto.

[0046] In the present invention, the information providing method for determining the equal producer comprises: providing Christensenellaceae, Leuconostocaceae, Peptostreptococcaceae, Adlercreutzia, Catenibacillus, Christensenellaceae R7 group, Eubacterium coprostanoligenes group, Eubacterium ruminantium group, Family XIII UCG001, Intestinibacter, Leuconostoc, NK4A214 group, Roseburia, Ruminococcus and UCG010 from a sample of an individual. The method may further include, but is not limited to, a step of determining an individual as an equal producer when one or more intestinal microorganisms selected from the group are up-expressed in a sample of the individual compared to equal non-producers.

[0047] In the present invention, the method for providing information for determining an equal producer may further include, but is not limited to, a step of determining an individual as an equal producer when one or more intestinal microorganisms selected from the group consisting of Streptococcaceae, Blautia, Eggerthella, and Streptococcus are down-expressed in a sample of the individual compared to an equal non-producer.

[0048] In addition, the present invention relates to a method for treating Peptostreptococcaceae, Christensenellaceae, Streptococcaceae, Leuconostocaceae, Eggerthella, Adlercreutzia, Family XIII UCG001, Intestinibacter, Ruminococcus, NK4A214 group, UCG010, Christensenellaceae R7 group, Streptococcus, Eubacterium coprostanoligenes group, Eubacterium ruminantium group, Catenibacillus, Provided is a use for determining equal producers of a composition comprising, as an active ingredient, one or more intestinal microorganisms selected from the group consisting of Leuconostoc, Roseburia, and Blautia.

[0049] In addition, the present invention provides a method for producing an equal producer determination agent, comprising the steps of: producing a preparation for determining an equal producer, Peptostreptococcaceae, Christensenellaceae, Streptococcaceae, Leuconostocaceae, Eggerthella, Adlercreutzia, Family XIII UCG001, Intestinibacter, Ruminococcus, NK4A214 group, UCG010, Christensenellaceae R7 group, Streptococcus, Eubacterium coprostanoligenes group, Eubacterium ruminantium group, Provided is a use of a composition comprising, as an active ingredient, one or more intestinal microorganisms selected from the group consisting of Catenibacillus, Leuconostoc, Roseburia, and Blautia.

[0050] The intestinal microbial biomarker according to the present invention shows different expression levels in equal producers and non-producers, so that it is possible to easily determine whether a person is an equal producer without going through a clinical trial process simply by confirming the expression level of the biomarker.

[0051] Therefore, it is expected that the present invention will contribute to the development of personalized health management and disease prevention strategies, and furthermore, help create a healthy intestinal microbiome environment.

[0052] Figure 1 is a diagram showing the results of a comparative analysis of alpha diversity of samples before (V2) and after (V3) consumption of soy isoflavone health functional food between equal producers and non-producers.

[0053] Figure 2 is a diagram showing the results of a comparative analysis (Unweighted / Weighted method) of beta-diversity of samples before (V2) and after (V3) consumption of soy isoflavone health functional food between equal producers and non-producers.

[0054] Figure 3 is a diagram showing the results of a comparative analysis (Bray-Curtis / Jaccard method) of the beta diversity of samples before (V2) and after (V3) consuming soy isoflavone health functional food between equal producers and non-producers.

[0055] Figure 4 is a diagram showing the ROC-AUC curve for the performance of gut microbiota biomarkers selected from the equal generator.

[0056] Figures 5a and 5b are diagrams showing the box-plot results of the gut microbiota biomarkers for discrimination of equal producers / non-producers selected through the Boruta technique.

[0057] Below, with reference to the attached drawings, embodiments of the present invention are described in detail so that those skilled in the art can easily implement them. However, the present invention may be implemented in various different forms and is not limited to the embodiments described herein.

[0058] Throughout this specification, when a part is said to "include" a certain component, this does not exclude other components, but rather includes other components, unless otherwise specifically stated. As used throughout this specification, the terms "step of" or "step of" do not mean "step for."

[0059] Throughout the present 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 the components.

[0060] [Example]

[0061] Example 1. Analysis of differences in gut microbiota communities between equal producers and non-producers (NGS analysis)

[0062] The gut microbiota was extracted from fecal samples according to the manufacturer's manual, and DNA concentration was measured using a fluorescent marker. Libraries were then constructed and sequenced using sequencing equipment (Miseq, Illumina). Metagenomic analysis, such as next-generation sequencing, was used to analyze the gut microbiota differences between equal-producers and non-producers, using various techniques, including comparison of gut microbiota diversity.

[0063] Example 1.1. Results of comparative analysis of alpha diversity of samples before (V2) and after (V3) consumption of soy isoflavone health functional food between equal producers and non-producers

[0064] To compare changes in gut microbial diversity across sample groups, alpha diversity indices were analyzed. Alpha diversity indices include Observed, Chao1, Shannon, and Simpson.

[0065] Alpha diversity is divided into a Richness index, which increases in value as the sample contains more species, and an Evenness index, which considers even the ratio. Observed and Chao1 are Richness indexes, while Shannon and Simpson are Evenness indexes.

[0066] For the Richness metric, the value increases as the sample contains more diverse species. Observed is the simplest metric, representing the number of microbial species present, providing an intuitive insight into the diversity of microbial species present in each sample. Chao1 is a richness-based metric, calculated by taking into account even the presence of rare species.

[0067] The Evenness index increases as the sample distribution becomes more uniform. Shannon's index reaches its maximum value when all species have the same proportion. Simpson's index, when two randomly selected sequences are chosen, represents the probability that the two sequences are of the same species, and has a value between 0 and 1.

[0068] Accordingly, the inventors divided the sample group into equal producers / non-producers and compared and analyzed the values ​​of various alpha diversity indices (Observed, Chao1, Shannon, Simpson) of the same subjects before (V2) and after (V3) ingestion of soy isoflavones. The results are shown in Fig. 1.

[0069] As shown in Figure 1, when comparing before (V2) or after (V3) soy isoflavone intake, equal producers had significantly higher alpha diversity than equal non-producers in most cases (p<0.05). However, when comparing before (V2) and after (V3) soy isoflavone intake in each sample group of equal producers and non-producers, there was no significant change in alpha diversity.

[0070] That is, the Richness and Evenness indices of equal producers are higher than those of equal non-producers, which means that equal producers contain more diverse microbial species and that the distribution among microbial species is more uniform than that of equal non-producers. In addition, there was no significant change in alpha diversity when comparing the sample groups of equal producers and non-producers before (V2) and after (V3) ingestion of soy isoflavones, indicating that the microbial community within each sample was not significantly affected by one week of soy isoflavones ingestion.

[0071] Example 1.2. Comparative analysis of beta diversity of samples before (V2) and after (V3) consumption of soy isoflavone health functional food between equal producers and non-producers.

[0072] Beta diversity is an analytical method used to compare the diversity of microbial communities in two or more samples.

[0073] Unweighted is calculating the distance using the presence or absence of microbial flora, and Weighted is calculating the distance using the amount of microbial flora so that it can be viewed visually.

[0074] Bray-Curtis is a method that calculates based on the abundance of species, while Jaccard is a method that calculates the presence or absence of species between samples and does not consider abundance.

[0075] Accordingly, the present inventors compared and analyzed the beta diversity of samples before (V2) and after (V3) ingestion of soy isoflavone health functional foods between equal producers and non-producers. The results are shown in Figures 2 and 3.

[0076] As shown in Figures 2 and 3, the p-values ​​of Unweighted, Weighted, Bray-Curtis, and Jaccard were 0.002, 0.032, 0.001, and 0.001, respectively, indicating that the beta diversity difference between equal generators and non-generators was significant.

[0077] Thus, the microbial communities of equal producers and equal non-producers showed significant differences in beta diversity, and the inventors confirmed the possibility of using specific microorganisms as biomarkers for identifying equal producers.

[0078] Example 2. Method for selecting gut microbiota biomarkers using an equal generator / non-generator discrimination algorithm.

[0079] To select gut microbiota biomarkers that can distinguish equal producers / non-producers, the Boruta technique was applied, and feature importance based on random forests was calculated to select valid gut microbiota.

[0080] To confirm the performance of the gut microbiota biomarkers selected from the equal generator, the AUC value for the ROC curve was calculated and evaluated.

[0081] The ROC-AUC curve (Receiver Operating Characteristic - Area Under the Curve) is a tool for evaluating the performance of binary classification models. It visualizes the relationship between a model's True Positive Rate (TPR) and False Positive Rate (FPR) based on various thresholds. This curve allows for a visual assessment of the predictive performance of a particular model.

[0082] The AUC value represents the area under the ROC curve and represents the overall performance of the model as a single number. An AUC of 0.5 indicates a model that makes random predictions. A value of 0.5 < AUC < 1 indicates a model with good performance, and an AUC of 1 indicates a model with perfect predictions. Similarly, if the AUC value is close to 1 and the model fit (p value) is lower than 0.05, the model has good predictive performance in identifying equal generators. The results are shown in Figure 4.

[0083] As shown in Figure 4, the model was confirmed to be a significant model with very high predictive power, with an AUC of 0.9796, close to 1, and a p value of 0.002306.

[0084] Example 3. Box plot analysis results of gut microbiota biomarkers for identifying equal producers / non-producers selected using the Boruta technique.

[0085] The gut microbiota biomarkers for discrimination of equal producers / non-producers selected through the Boruta technique were Peptostreptococcaceae, Christensenellaceae, Streptococcaceae, and Leuconostocaceae at the family level, and Eggerthella, Adlercreutzia, Family XIII UCG001, Intestinibacter, Ruminococcus, NK4A214 group, UCG010, Christensenellaceae R7 group, Streptococcus, and Eubacterium coprostanoligenes at the genus level. The genera selected were Eubacterium coprostanoligenes group, Eubacterium ruminantium group, Catenibacillus, Leuconostoc, Roseburia, and Blautia.

[0086] The box plot results of the gut microbiota biomarkers for discrimination of equal producers / non-producers selected through the above-mentioned Boruta technique are shown in Figures 5a and 5b.

[0087] As shown in Figures 5a and 5b, Christensenellaceae, Leuconostocaceae, Peptostreptococcaceae, Adlercreutzia, Catenibacillus, Christensenellaceae R7 group, Eubacterium coprostanoligenes group, Eubacterium ruminantium group, Family XIII UCG001, Intestinibacter, Leuconostoc, NK4A214 group, Roseburia, Ruminococcus, and UCG010 were found to be equal in the equal producer. It showed significantly higher results compared to non-producers.

[0088] Additionally, Streptococcaceae, Blautia, Eggerthella, and Streptococcus showed significantly higher results in equal non-producers compared to equal producers.

[0089] The present inventors analyzed the differences in gut microbial communities between equal producers and non-producers by comparing alpha and beta diversity, and selected gut microbial biomarkers for identifying equal producers using the Boruta technique. Therefore, it is expected that the gut microbial biomarkers of the present invention for identifying equal producers will be able to easily distinguish equal producers from non-producers.

Claims

1. Peptostreptococcaceae, Christensenellaceae, Streptococcaceae, Leuconostocaceae, Eggerthella, Adlercreutzia, Family XIII UCG001, Intestinibacter, Ruminococcus, NK4A214 group, UCG010, Christensenellaceae R7 group, Streptococcus, Eubacterium coprostanoligenes group, Eubacterium ruminantium group, Catenibacillus, An intestinal microbial biomarker composition for determining equal producers, comprising as an active ingredient one or more intestinal microorganisms selected from the group consisting of Leuconostoc, Roseburia, and Blautia.

2. In paragraph 1, At least one intestinal microorganism selected from the group consisting of Christensenellaceae, Leuconostocaceae, Peptostreptococcaceae, Adlercreutzia, Catenibacillus, Christensenellaceae R7 group, Eubacterium coprostanoligenes group, Eubacterium ruminantium group, Family XIII UCG001, Intestinibacter, Leuconostoc, NK4A214 group, Roseburia, Ruminococcus, and UCG010 is compared to an equal non-producer A composition of intestinal microorganism biomarkers for determining equal producers, characterized by upward expression in the producer.

3. In paragraph 1, An intestinal microbial biomarker composition for determining equal producers, characterized in that one or more intestinal microorganisms selected from the group consisting of Streptococcaceae, Blautia, Eggerthella, and Streptococcus are down-expressed in equal producers compared to equal non-producers.

4. Peptostreptococcaceae, Christensenellaceae, Streptococcaceae, Leuconostocaceae, Eggerthella, Adlercreutzia, Family XIII UCG001, Intestinibacter, Ruminococcus, NK4A214 group, UCG010, Christensenellaceae R7 group, Streptococcus, Eubacterium coprostanoligenes group, Eubacterium ruminantium group, Catenibacillus, A composition for determining equal producers, characterized in that it comprises a preparation capable of detecting one or more intestinal microorganisms selected from the group consisting of Leuconostoc, Roseburia, and Blautia.

5. In paragraph 4, A composition for determining equal producers, characterized in that the agent capable of detecting the intestinal microorganism is at least one selected from the group consisting of a primer, a probe, an aptamer, an avidity multimer, a peptidomimetic, and an antibody.

6. A kit for determining an equal generator, comprising the composition of Article 4.

7. In paragraph 6, A kit for determining an equal producer, characterized in that the above kit is an RT-PCR (Reverse transcription polymerase chain reaction) kit, a DNA chip kit, an ELISA (Enzymelinked immunosorbent assay) kit, a protein chip kit, a rapid kit, or an MRM (Multiple reaction monitoring) kit.

8. From the sample of the individual, Peptostreptococcaceae, Christensenellaceae, Streptococcaceae, Leuconostocaceae, Eggerthella, Adlercreutzia, Family XIII UCG001, Intestinibacter, Ruminococcus, NK4A214 group, UCG010, Christensenellaceae R7 group, Streptococcus, Eubacterium coprostanoligenes group, Eubacterium ruminantium group, A step of detecting one or more intestinal microorganisms selected from the group consisting of Catenibacillus, Leuconostoc, Roseburia, and Blautia; and A method for providing information for determining an equal producer, comprising: a step of comparing the expression level of the intestinal microorganisms with that of a control group.

9. In paragraph 8, At least one intestinal microorganism selected from the group consisting of Christensenellaceae, Leuconostocaceae, Peptostreptococcaceae, Adlercreutzia, Catenibacillus, Christensenellaceae R7 group, Eubacterium coprostanoligenes group, Eubacterium ruminantium group, Family XIII UCG001, Intestinibacter, Leuconostoc, NK4A214 group, Roseburia, Ruminococcus and UCG010 is isolated from a sample of an individual. A method for providing information for determining an equal generator, further comprising: a step of determining an entity as an equal generator when the entity is upwardly expressed in a sample of the entity compared to a non-generator; 10. In paragraph 8, A method for providing information for determining an equal producer, further comprising: a step of determining an individual as an equal producer when one or more intestinal microorganisms selected from the group consisting of Streptococcaceae, Blautia, Eggerthella, and Streptococcus are down-expressed in a sample of the individual compared to an equal non-producer;

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