Faecalibacterium longum and application thereof

Abstract

Provided are Faecalibacterium longum and applications thereof, in particular, the use of Faecalibacterium longum in the treatment and prevention of diabetes and related diseases, and a composition for treating and preventing diabetes and related diseases, including a pharmaceutical product, a beverage, a food product, or an animal feed composition, and the like, and a method for reducing body weight, fasting blood glucose, and / or blood lipids, and improving glucose tolerance.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of microbiology, and in particular, the present application relates to the use of Faecalibacterium longum in the treatment and prevention of diabetes and its related diseases, and also relates to compositions comprising Faecalibacterium longum and the use thereof. BACKGROUND

[0002] There are a large number of symbiotic microorganisms in the human intestinal tract, and these symbiotic microorganism populations constitute the "second organ" of the human body, which not only cooperates with the host to participate in the digestion and absorption of nutrients, but also plays an important role in maintaining the health of the human body. In terms of cell number, the number of all cells of intestinal microorganisms is 10 times the number of human cells, and these large microbial populations are mainly composed of two categories: Firmicutes and Bacteroidetes. More and more studies have shown that the composition and diversity of intestinal microorganisms are closely related to the occurrence of many diseases, such as obesity, diabetes, irritable bowel syndrome, ulcerative colitis, colon cancer, fatty liver, etc. Due to the influence of environment, diet, drugs, etc., the balance of intestinal microorganisms under the original healthy state is broken, leading to intestinal function disorder, and further inducing the occurrence of the above several diseases.

[0003] In recent years, with the improvement of people's daily life level, the dietary structure is gradually unbalanced, and the incidence of metabolic diseases such as obesity and diabetes is rapidly increasing. According to the statistics of the World Health Organization, about 422 million people worldwide suffered from diabetes in 2014, and the number of deaths caused by diabetes reached 1.5 million by 2012. Therefore, diabetes has become a major public health problem at present. Diabetes is a disease caused by abnormally high blood sugar, which is mainly a metabolic disorder caused by insufficient insulin secretion or insulin function defects. Common high-risk factors for the occurrence of diabetes include genetic factors, environmental factors and unhealthy living habits. The occurrence of diabetes is accompanied by the occurrence of many complications, such as coronary heart disease, hypertension, myocardial infarction, stroke, senile dementia, Parkinson's disease, kidney disease, retinopathy, etc., which seriously reduce the life quality of patients. Among them, type 2 diabetes is a non-insulin-dependent diabetes, and the number of patients accounts for 90% of all diabetes patients. A large number of studies have shown that the incidence of type 2 diabetes is closely related to the disorder of intestinal flora. Intestinal flora is closely related to the regulation of energy balance and inflammatory response between the host, and the incidence of type 2 diabetes is mainly caused by insulin resistance and low-level inflammatory response.

[0004] The hypoglycemic drugs currently used for treating diabetes include sulfonylureas, α-glucosidase inhibitors and biguanides. Sulfonylureas mainly promote insulin secretion, but can cause serious liver and kidney damage and are not suitable for people with allergies. α-glucosidase inhibitors mainly inhibit the activity of α-amylase and intestinal α-glucosidase to inhibit the hydrolysis of carbohydrates, thereby reducing postprandial blood glucose, but can easily cause adverse reactions such as abdominal distension and diarrhea. Biguanides can achieve the purpose of reducing blood sugar by regulating blood sugar transport, such as delaying sugar absorption, promoting glucose decomposition, inhibiting liver glucose production, and increasing glucose transfer protein. Biguanides can also cause some side effects, such as gastrointestinal discomfort, diarrhea, vomiting, skin rash, and inactivation after long-term use.

[0005] Therefore, there is an urgent need in the art to develop a new drug with no toxic side effects for treating and preventing diabetes and related diseases. SUMMARY

[0006] Another object of the present application is to provide the use of Faecalibacterium longum in treating and preventing diabetes and related diseases.

[0007] Another object of the present application is to provide an effective drug, beverage, food composition, or animal feed composition with no toxic side effects for treating and preventing diabetes and related diseases.

[0008] Another object of the present application is to provide a method for reducing body weight, fasting blood glucose and / or blood lipids and its application.

[0009] Another object of the present application is to provide a method for improving sugar tolerance and its application.

[0010] The first aspect of the present application provides a Faecalibacterium longum, which is Faecalibacterium longum.

[0011] In another preferred embodiment, the sequence of 16s rDNA of the Faecalibacterium longum is shown in SEQ ID NO. 1.

[0012] In another preferred embodiment, the Faecalibacterium longum is Faecalibacterium longum CM04-06, with the preservation number of CGMCC 1.5208.

[0013] The second aspect of the present application provides a composition comprising: (a) a safe and effective amount of the Faecalibacterium longum of claim 1 and / or its metabolites; and (b) a food acceptable carrier or a pharmaceutically acceptable carrier.

[0014] In another preferred embodiment, the composition further comprises a milk growth factor.

[0015] In another preferred embodiment, the composition is selected from the group consisting of a food composition, a health composition, a pharmaceutical composition, a beverage composition, a feed composition, or a combination thereof.

[0016] In another preferred embodiment, the composition is an oral formulation.

[0017] In another preferred embodiment, the composition is a liquid formulation, a solid formulation, or a semi-solid formulation.

[0018] In another preferred embodiment, the composition is in a dosage form selected from the group consisting of a powder, a granule, a tablet, a sugar-coated tablet, a capsule, a granule, a suspension, a solution, a syrup, a drop, and a sublingual tablet.

[0019] In another preferred embodiment, the food composition comprises a milk product, a solution product, a powder product, or a suspension product.

[0020] In another preferred embodiment, the food composition comprises a milk product, a milk powder, or a milk emulsion.

[0021] In another preferred embodiment, the liquid formulation is selected from the group consisting of a solution product or a suspension product.

[0022] In another preferred embodiment, the composition contains 1 x 10 10 cfu / mL or cfu / g of Faecalibacterium longum CM04-06, preferably 1 x 10 4 -1 x 10 10 cfu / mL or cfu / g of Faecalibacterium longum CM04-06, based on the total volume or the total weight of the composition.

[0023] In another preferred embodiment, the composition contains 0.0001-99 wt%, preferably 0.1-90 wt% (check) of the Faecalibacterium longum and / or the metabolite thereof, based on the total weight of the composition.

[0024] In another preferred embodiment, the composition is in a unit dosage form (one tablet, one capsule, or one vial), and the mass of the composition in each unit dosage form is 0.05-5 g, preferably 0.1-1 g.

[0025] In another preferred embodiment, the composition further comprises other probiotics and / or prebiotics.

[0026] In another preferred embodiment, the probiotic is selected from the group consisting of a lactic acid bacterium, a Bifidobacterium, a Lactobacillus acidophilus, or a combination thereof.

[0027] In another preferred embodiment, the prebiotic is selected from the group consisting of fructo-oligosaccharides (FOS), galacto-oligosaccharides (GOS), xylo-oligosaccharides (XOS), lactulose (LACT), soybean oligosaccharides (SOS), inulin, or a combination thereof.

[0028] The third aspect of the present application provides use of the Faecalibacterium prausnitzii of the first aspect of the present application, or the composition of the second aspect of the present application, in the manufacture of a medicament or a preparation for one or more uses selected from the group consisting of: (a) preventing and / or treating obesity; (b) reducing blood lipid; (c) preventing or treating cardiovascular disease; and / or (d) preventing and / or treating diabetes.

[0029] In another preferred embodiment, the preparation comprises a microecological preparation.

[0030] The fourth aspect of the present application provides use of the Faecalibacterium prausnitzii of the first aspect of the present application, or the composition of the second aspect of the present application, in the manufacture of a medicament or a preparation for one or more uses selected from the group consisting of:

[0031] (i) inhibiting weight gain in a mammal;

[0032] (ii) reducing blood lipid level in a mammal;

[0033] (iii) increasing high-density lipoprotein (HDL-C) level in a mammal;

[0034] (iv) reducing low-density lipoprotein (LDL-C) level in a mammal;

[0035] (v) reducing blood glucose level in a mammal;

[0036] (vi) improving glucose tolerance in a mammal.

[0037] In another preferred embodiment, the mammal comprises a human, a rodent (e.g. a rat, a mouse).

[0038] In another preferred embodiment, the reducing blood lipid level in a mammal comprises reducing total cholesterol (TC) level and / or triglyceride level.

[0039] In another preferred embodiment, the reducing blood glucose level in a mammal comprises reducing fasting blood glucose level.

[0040] The fifth aspect of the present application provides a method for preparing the composition of the second aspect of the present application, comprising the steps of:

[0041] The Faecalibacterium prausnitzii and / or metabolite thereof of the first aspect of the application is mixed with a food acceptable carrier or a pharmaceutically acceptable carrier to form the composition of the second aspect of the application.

[0042] In another preferred embodiment, the method further comprises the step of mixing the Faecalibacterium prausnitzii and / or metabolite thereof with a growth factor.

[0043] In another preferred embodiment, the growth factor is a milk growth factor.

[0044] In another preferred embodiment, the growth factor is selected from the group consisting of a vitamin, a purine, a pyrimidine, or a combination thereof.

[0045] In another preferred embodiment, the composition is an oral formulation.

[0046] The sixth aspect of the application provides a method of production comprising the steps of:

[0047] (a) culturing the Faecalibacterium prausnitzii of the first aspect of the application under conditions suitable for growth to obtain a culture product;

[0048] (b) optionally, isolating the Faecalibacterium prausnitzii and / or metabolite thereof from the culture product; and

[0049] (c) optionally, mixing the Faecalibacterium prausnitzii and / or metabolite thereof isolated from the previous step with a food acceptable carrier or a pharmaceutically acceptable carrier to form a composition.

[0050] In another preferred embodiment, prior to step (c), the method further comprises the step of mixing the Faecalibacterium prausnitzii and / or metabolite thereof isolated from the previous step with a growth factor.

[0051] In another preferred embodiment, the growth factor is a milk growth factor.

[0052] In another preferred embodiment, the growth factor is selected from the group consisting of a vitamin, a purine, a pyrimidine, or a combination thereof.

[0053] The seventh aspect of the application provides a method of reducing body weight, fasting blood glucose and / or blood lipids by administering to the subject the Faecalibacterium prausnitzii of the first aspect of the application and / or metabolite thereof, or the composition of the second aspect of the application.

[0054] In another preferred embodiment, the administration is oral.

[0055] In another preferred embodiment, the administration is at a dose of 0.01-5 g per 50 kg body weight per day, preferably 0.1-2 g per 50 kg body weight per day.

[0056] In another preferred embodiment, the subject includes a mammal, such as a human.

[0057] The eighth aspect of the present application provides a method for improving glucose tolerance, administering the Faecalibacterium longum and / or its metabolite according to the first aspect of the present application, or the composition according to the second aspect of the present application to the subject.

[0058] In another preferred embodiment, the administration includes oral administration.

[0059] In another preferred embodiment, the administration dosage is 0.01-5 g / 50 kg body weight / day, preferably 0.1-2 g / 50 kg body weight / day.

[0060] In another preferred embodiment, the subject includes a mammal, such as a human.

[0061] It should be understood that, within the scope of the present application, each of the technical features of the present application described above and each of the technical features specifically described hereinafter (e.g. in the examples) can be combined with each other to form new or preferred technical solutions. Due to the limited space, they will not be listed one by one here. BRIEF DESCRIPTION OF DRAWINGS

[0062] Figure 1 Pictures showing colonies of Faecalibacterium longum cultured for 3 days.

[0063] Figure 2 Pictures showing gram staining of Faecalibacterium longum under a microscope (1000 times).

[0064] Figure 3 Pictures showing the changes in body weight of diabetic mice in the model group and treated with Faecalibacterium longum CM04-06 and metformin.

[0065] Figure 4 Pictures showing the results of fasting blood glucose (FBG) of diabetic mice in the model group and treated with Faecalibacterium longum CM04-06 and metformin.

[0066] Figure 5 Pictures showing the glucose tolerance of diabetic mice in the model group and treated with Faecalibacterium longum CM04-06 and metformin. DETAILED DESCRIPTION

[0067] The present inventors have made extensive and in-depth research and experiments, and unexpectedly found that Faecalibacterium longum has the effect of preventing and treating diabetes and related diseases (such as cardiovascular diseases and obesity diseases), and feeding an active composition containing Faecalibacterium longum to a test subject, it is found that the composition can inhibit weight gain, reduce blood lipids, reduce fasting blood glucose, improve sugar tolerance, and effectively alleviate diabetes, cardiovascular and obesity diseases. On this basis, the present application is completed.

[0068] As used herein, the term "comprising" means that various components can be used together in a mixture or composition of the present application. Therefore, the terms "consisting essentially of and "consisting of are included in the term "comprising".

[0069] As used herein, the terms "growth factor", "milk growth factor" are used interchangeably and include nutritional substances such as vitamin substances, purine substances, pyrimidine substances, or combinations thereof.

[0070] The vitamin substances include (but are not limited to) vitamin C, vitamin E, vitamin A, vitamin A precursors, vitamin B6, vitamin D3, vitamin K, folic acid, or combinations thereof.

[0071] The purine substances include (but are not limited to) purine nucleosides, wherein the purine nucleosides include 5'-phosphates of purine nucleosides selected from the group consisting of inosinic acid (inosine-5'-phosphate; IMP), guanylic acid (guanosine-5'-phosphate; GMP), xanthic acid (xanthine nucleoside-5'-phosphate; XMP), adenosine acid (adenosine-5'-phosphate; AMP), or combinations thereof.

[0072] The pyrimidine substances include all substances containing pyrimidine structures.

[0073] Faecalibacterium longum and its application

[0074] As used herein, the terms “faecalibacterium longum” and “faecalibacterium longum” are used interchangeably. In a preferred embodiment, the strain is Faecalibacterium longum CM04-06, accession number CGMCC 1.5208, isolated from human feces. The physiological characteristics of *Faecalibacterium longiflorum* are as follows: When cultured in an anaerobic environment at 37°C for 2-3 days, *Faecalibacterium longiflorum* colonies are yellowish-white, with high water content, slightly viscous, nearly round, opaque, flat with a raised center, and approximately 2-3 mm in diameter. Gram staining and microscopic observation show that CM04-06 cells are rod-shaped, Gram staining is negative, and no spores or flagella were found. The cell diameter is approximately 1 μm, and the length is 4-10 μm. Furthermore, the *Faecalibacterium longiflorum* described in this invention exhibits negative oxidase and catalase reactions. Its growth temperature range is 30-45°C, and its pH range is 4.0-9.0, with the optimal temperature and pH being 37°C and pH 7.0, respectively. It can tolerate 3% NaCl. It can ferment several carbohydrates, including mannose, raffinose, and trehalose, mainly producing formic acid, acetic acid, butyric acid, and lactic acid. In addition, it can also ferment to produce extracellular polysaccharides.

[0075] This invention provides the application of *Faecalibacterium longum* in the treatment and prevention of diabetes and related diseases (such as cardiovascular disease and obesity). When subjects consume a high-fat diet, the strain *Faecalibacterium longum* CM04-06 has one or more uses selected from the group consisting of: (i) inhibiting weight gain in the subject; (ii) lowering blood lipids; (iii) lowering fasting blood glucose levels; and (iv) improving glucose tolerance. According to a preferred embodiment of the invention, C57bL / 6 mice were used as experimental mice, fed a high-fat diet, and induced with streptourea (STZ) to obtain type 2 diabetes (T2D) model mice. T2D model mice treated with strain *Faecalibacterium longum* CM04-06, compared with the untreated control group (model group), showed slower weight gain, lower blood lipids, and decreased levels of various diabetes-related indicators, such as fasting blood glucose levels. Furthermore, glucose tolerance was significantly improved. Therefore, the strain can be used to prevent and treat diabetes and related diseases (such as cardiovascular disease and obesity).

[0076] Compositions and their applications

[0077] The present application also provides a composition, preferably a pharmaceutical composition. The composition comprises an effective amount of Faecalibacterium prausnitzii, and in a preferred embodiment, the composition further comprises a milk growth factor. In a preferred embodiment, the composition further comprises a probiotic selected from the group consisting of a lactic acid bacteria, a bifidobacteria, a lactobacillus acidophilus, or a combination thereof; and / or a prebiotic selected from the group consisting of fructo-oligosaccharides (FOS), galacto-oligosaccharides (GOS), xylo-oligosaccharides (XOS), lactulose (LACT), soybean oligosaccharides (SOS), inulin, or a combination thereof.

[0078] In a preferred embodiment, the composition is a liquid formulation, a solid formulation, or a semi-solid formulation.

[0079] In a preferred embodiment, the liquid formulation is selected from the group consisting of a solution formulation or a suspension formulation.

[0080] In a preferred embodiment, the dosage form of the composition is selected from the group consisting of a powder, a dust, a tablet, a sugar-coated tablet, a capsule, a granule, a suspension, a solution, a syrup, a drop, and a sublingual tablet.

[0081] The pharmaceutical composition of the present application can be administered in any form of a pharmaceutical tablet, a needle, or a capsule, and the pharmaceutical formulation includes excipients, pharmaceutically acceptable vehicles and carriers, which can be selected according to the administration route. The pharmaceutical formulation of the present application can further comprise an auxiliary active ingredient.

[0082] Lactose, glucose, sucrose, sorbitol, mannose, starch, gum arabic, calcium phosphate, alginate, gelatin, calcium silicate, fine crystalline cellulose, polyvinylpyrrolidone (PVP), cellulose, water, sugar syrup, methyl cellulose, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, or mineral oil, etc. can be used as a carrier, an excipient, or a diluent, etc. for the pharmaceutical composition of the present application.

[0083] In addition, the pharmaceutical composition of the present application can further comprise a lubricant, a wetting agent, an emulsifier, a suspension stabilizer, a preservative, a sweetener, and a flavoring agent, etc. The pharmaceutical composition of the present application can be produced in an enteric formulation by various well-known methods, so that the active ingredient of the pharmaceutical composition, i.e. the microorganism, can smoothly pass through the stomach without being destroyed by gastric acid.

[0084] In addition, the microorganism of the present application can be used in the form of a capsule prepared by a conventional method. For example, a standard excipient and the freeze-dried microorganism of the present application are mixed to prepare a small pellet, and then the pellet is filled into a gelatin capsule. In addition, the microorganism of the present application and a pharmaceutically acceptable excipient such as a liquid gum, cellulose, silicate, or mineral oil, etc. are mixed to prepare a suspension or a dispersion, and the suspension or the dispersion can be filled into a soft gelatin capsule.

[0085] The pharmaceutical composition of the present application can be prepared in the form of enteric coated tablets for oral administration. The term "enteric coating" as used herein includes all conventional pharmaceutical acceptable coatings which are not degraded by gastric acid but are sufficiently broken down in the small intestine to release the microorganism of the present application rapidly. The enteric coating of the present application is capable of maintaining its integrity in a simulated gastric acid, e.g. HCl solution at pH 1 at 36-38°C for more than 2 hours and is preferably broken down in a simulated intestinal fluid, e.g. a buffer solution at pH 7.0 in 1.0 hour.

[0086] The enteric coating of the present application is coated at a level of about 16-30 mg per tablet, preferably 16-25 mg, more preferably 16-20 mg. The enteric coating of the present application has a thickness of 5-100 μm, preferably 20-80 μm. The enteric coating composition is selected from conventional polymers known in the art.

[0087] The preferred enteric coating of the present application is prepared from a copolymer of cellulose acetate phthalate polymer or cellulose acetate trimellitate polymer and methacrylic acid (e.g. a copolymer of methacrylic acid containing more than 40% methacrylic acid and a copolymer of methacrylic acid containing hydroxypropyl cellulose acetate phthalate or ester derivatives thereof).

[0088] The cellulose acetate phthalate used in the enteric coating of the present application has a viscosity of about 45-90 cps, an acetyl content of 17-26% and a phthalic acid content of 30-40%. The cellulose acetate trimellitate used in the enteric coating has a viscosity of about 5-21 cps and an acetyl content of 17-26%. Cellulose acetate trimellitate is manufactured by Eastman Kodak and can be used as the enteric coating material in the present application.

[0089] The hydroxypropyl methyl cellulose phthalate used in the enteric coating of the present application has a molecular weight of about 20,000-130,000 daltons, preferably 80,000-100,000 daltons, a hydroxypropyl content of 5-10%, a methoxyl content of 18-24% and a phthaloyl content of 21-35%.

[0090] The hydroxypropyl methyl cellulose phthalate used in the enteric coating of the present application is HP50 manufactured by Shin-Etsu Chemical Co. Ltd. of Japan. HP50 has a hydroxypropyl content of 6-10%, a methoxyl content of 20-24% and a phthaloyl content of 21-27% and a molecular weight of 84,000 daltons. Another enteric coating material is HP55 which has a hydroxypropyl methyl cellulose phthalate content of 5-9%, a methoxyl content of 18-22% and a phthaloyl content of 27-35% and a molecular weight of 78,000 daltons.

[0091] The enteric coating is prepared by spraying the enteric coating solution onto the core using conventional methods. All solvents in the enteric coating process are alcohols (e.g., ethanol), ketones (e.g., acetone), halogenated hydrocarbon compounds (e.g., methylene chloride), or combinations thereof. Softening agents such as di-n-butyl phthalate and triacetin are added to the enteric coating solution in a ratio of about 1 part of the coating material to about 0.05 parts or about 0.3 parts of the softening agent. The spraying process is preferably carried out continuously, and the amount of material sprayed can be controlled according to the conditions used for the coating. The spraying pressure can be adjusted at will, and in general, satisfactory results are obtained at an average pressure of 1 to 1.5 bar.

[0092] "Pharmaceutically effective amount" in the specification means an amount that is functional or active in humans and / or animals and is acceptable to humans and / or animals. For example, in the present application, a preparation containing 1 x 10 10 cfu / ml or cfu / g (in particular, it can contain 1 x 10 4 -1 x 10 10 cfu / ml or cfu / g; more particularly, it can contain 1 x 10 6 -1 x 10 10 cfu / ml or cfu / g) of Faecalibacterium prausnitzii and / or its metabolite can be prepared.

[0093] When used for the preparation of a pharmaceutical composition, the effective dose of Faecalibacterium prausnitzii or its metabolite used can vary depending on the mode of administration and the severity of the disease to be treated. The dosage form suitable for oral administration contains about 1 x 10 10 cfu / ml or cfu / g (in particular, it can contain 1 x 10 4 -1 x 10 10 cfu / ml or cfu / g; more particularly, it can contain 1 x 10 6 -1 x 10 10 cfu / ml or cfu / g) of active Faecalibacterium prausnitzii or active ingredients produced by fermentation. This dosage regimen can be adjusted to provide an optimal therapeutic response. For example, several divided doses can be administered daily, or the dose can be proportionally reduced, as required by the exigencies of the therapeutic situation.

[0094] The Faecalibacterium prausnitzii or metabolite thereof can be administered by oral or other routes. Solid carriers include starch, lactose, dicalcium phosphate, microcrystalline cellulose, sucrose, and kaolin, while liquid carriers include culture media, polyethylene glycol, non-ionic surfactants, and edible oils such as corn, peanut, and sesame oils. Adjuvants commonly used in preparing pharmaceutical compositions can also be advantageously included, such as flavoring agents, coloring agents, preservatives, and antioxidants such as vitamin E, vitamin C, BHT, and BHA.

[0095] From the standpoint of ease of preparation and administration, the preferred pharmaceutical composition is a solid composition, especially tablets and hard or liquid filled capsules. Oral administration is preferred.

[0096] The composition of the present application is administered to the individual one or more times per day. The dosage unit is that form in which the active ingredient is contained in a pharmaceutically acceptable carrier and is appropriate for the patient to be treated, whether human or other mammal. Each unit contains a therapeutically effective amount of the microorganism of the present application. The amount administered varies depending on the weight of the patient, the severity of the diabetes, the supplementary active ingredients included, and the microorganism used. Furthermore, administration can be separated, if possible, and continuous, if necessary. Thus, the amount administered does not limit the present application. Furthermore, the "composition" in the present application means not only a pharmaceutical product but also a functional food and a health supplement food. In a preferred embodiment, the composition includes a beverage, a food, a pharmaceutical product, an animal feed, etc.

[0097] In a preferred embodiment of the present application, there is also provided a food composition comprising an effective amount of Faecalibacterium prausnitzii and / or a metabolite thereof, and a remainder of a food acceptable carrier, the food composition being in a dosage form selected from a solid, a dairy, a solution, a powder, or a suspension. In a preferred embodiment, the food composition can further comprise a milk growth factor.

[0098] In a preferred embodiment, the composition is formulated as follows:

[0099] 1 x 10 10 Faecalibacterium prausnitzii and / or a metabolite thereof at a concentration of 1 x 10

[0100] In another preferred embodiment, the composition is formulated as follows:

[0101] 1 x 10 6 -1 x 10 10 Faecalibacterium prausnitzii and / or a metabolite thereof at a concentration of 1 x 10

[0102] Microecological preparation

[0103] A microecological preparation is a biological preparation containing probiotics and metabolites or a dietary supplement that can increase probiotics, which can regulate and maintain the microecological balance in the intestinal tract, and achieve the purpose of improving the health level of the human body. It mainly includes probiotics, prebiotics and synbiotics.

[0104] In the present application, the microecological preparation comprises (a) a safe and effective amount of Faecalibacterium prausnitzii and / or its metabolites; and (b) a food-acceptable carrier or a pharmaceutically acceptable carrier. In a preferred embodiment, the preparation further comprises growth factors (such as milk growth factors, preferably including vitamin substances, purine substances, and / or pyrimidine substances).

[0105] Method for producing Faecalibacterium prausnitzii

[0106] Generally, Faecalibacterium prausnitzii can be prepared by conventional methods.

[0107] In the present application, a method for mass-producing Faecalibacterium prausnitzii is provided, which specifically comprises the following steps:

[0108] (a) culturing the Faecalibacterium prausnitzii under suitable conditions for cultivation, thereby obtaining a culture product;

[0109] (b) optionally, isolating Faecalibacterium prausnitzii cells and / or its metabolites from the culture product; and

[0110] (c) optionally, mixing the Faecalibacterium prausnitzii cells and / or its metabolites obtained by isolation in the previous step with a food-acceptable carrier or a pharmaceutically acceptable carrier, thereby preparing a composition.

[0111] Method for reducing body weight, fasting blood glucose and / or blood lipids

[0112] In another preferred embodiment, the method comprises taking the pharmaceutical composition, food composition, beverage composition, or combination thereof of the present application. The experimental subject is a human.

[0113] In another preferred embodiment, the method comprises taking the pharmaceutical composition, food composition, or animal feed of the present application, or combination thereof. The experimental subject is an animal, preferably a rodent, a rabbit.

[0114] Method for improving glucose tolerance

[0115] In another preferred embodiment, the method comprises taking the pharmaceutical composition, food composition, beverage composition, or combination thereof of the present application. The experimental subject is a human.

[0116] In another preferred embodiment, the method comprises ingesting the pharmaceutical composition, the food composition, or the animal feed of the present application, or a combination thereof. The subject is an animal, preferably a murine or a lagomorph.

[0117] Bacterial strain preservation

[0118] The bacterial strain Faecalibacterium longum CM04-06 (same as the deposit name) of the present application has been deposited at the China General Microbiological Culture Collection Center (CGMCC) on June 13, 2016, at No. 1, Yihuangyuan 3rd, Beichenxi Road, Chaoyang District, Beijing, China, with the accession number: CGMCC 1.5208.

[0119] The main advantages of the present application include:

[0120] (a) The Faecalibacterium longum of the present application can significantly reduce body weight, blood lipid, and fasting blood glucose level.

[0121] (b) The Faecalibacterium longum of the present application can significantly reduce the indicators (such as cholesterol and triglyceride) associated with obesity and its related diseases (such as cardiovascular disease).

[0122] (c) The Faecalibacterium longum of the present application can significantly reduce the levels of total cholesterol, triglyceride, and low-density lipoprotein.

[0123] (d) The Faecalibacterium longum of the present application can significantly increase the level of high-density lipoprotein.

[0124] (e) The Faecalibacterium longum of the present application can significantly improve the glucose tolerance.

[0125] The present application will be further described in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present application and not used to limit the scope of the present application. The experimental methods in the following examples, if not specified, are generally carried out according to the conventional conditions, such as the conditions described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or the conditions described in Microbiology: A Laboratory Manual (James Cappuccino and Natalie Sherman, Pearson Education Press), or the conditions recommended by the manufacturer.

[0126] Unless otherwise specified, the materials used in the examples are commercially available products.

[0127] Faecalibacterium prausnitzii ATCC 27768, hereinafter referred to as ATCC 27768, was purchased from American Type Culture Collection, and the preservation number is ATCC 27768.

[0128] Example 1 Screening and identification of Faecalibacterium prausnitzii CM04-06

[0129] 1.1 Isolation and culture of CM04-06

[0130] Faecalibacterium prausnitzii CM04-06, hereinafter referred to as CM04-06, was isolated from a stool sample of a healthy child (male) in Shenzhen. The isolation and culture environment was strictly anaerobic condition, and the specific isolation process was as follows: 0.2 g of stool sample was taken in an anaerobic operation box, suspended and dispersed with 1 ml of sterile PBS, mixed thoroughly after sufficient shaking, gradient diluted and plated, and the culture medium was anaerobic PYG medium (purchased from HuanKai Microbial Technology Co., Ltd.), the specific components were as follows:

[0131] Table 1

[0132] Component Content (1 L) Peptone 5g Trypsinized casein 5g Yeast powder 10g Beef extract 5g Glucose 5g K2HPO4 2g Sodium acetate 5g [Cysteine-HCl·H2O] 0.5g Hematin 5 mg Inorganic salt solution 40ml Resazurin 1 mg Distilled water 950ml

[0133] The formula of the inorganic salt solution is as follows:

[0134]

[0135]

[0136] The plated plate was cultured at 37°C under anaerobic condition for 3-4 days, after the colonies grew on the surface of the plate, single colonies were picked and streaked for isolation and purification until a pure culture strain was obtained, and the isolated strain was frozen and preserved at-80°C with glycerol.

[0137] 1.2 16S rDNA identification of Faecalibacterium prausnitzii CM04-06

[0138] (1) Genomic extraction: the isolated strain was cultured, and when the bacterial concentration reached 10 8 cfu / ml, 2 ml of bacterial solution was taken for genomic DNA extraction.

[0139] (2) PCR amplification of 16S rDNA: 16S rDNA was amplified using DNA as template, and the amplification primers were universal amplification primers of 16S rDNA: 8F-1492R (5'- AGAGTTTGATCATGGCTCAG-3' (SEQ ID NO.: 2) and 5'-TAGGGTTACCTTGTTACGACTT-3' (SEQ ID NO.: 3)), and the PCR amplification procedure was as follows:

[0140]

[0141]

[0142] (3) Purification and sequencing: the PCR product was purified by magnetic beads, and then electrophoresis detection was performed, and the band position of 16S rDNA was about 1.5k, and the purified product was subjected to 3730 sequencing.

[0143] (4) 16S rDNA sequence database comparison: 1372bp of 16S rDNA sequence was obtained by sequencing, and the sequence was compared in EzTaxon-e database, and the species classification information of the strain was obtained. According to the information of 16S rDNA, it can be preliminarily judged that CM04-06 belongs to a new species of Faecalibacterium genus.

[0144] The 16S rDNA sequence of CM04-06 is SEQ ID NO.: 1, as shown below:

[0145]

[0146] 1.3 Evolution analysis of 16S rDNA of CM04-06

[0147] The evolution analysis of CM04-06 used 16S rDNA sequence, and the EzTaxon-e database was compared by the 16S rDNA sequence of CM04-06, and the species with close genetic relationship with CM04-06 was obtained, and the 16S rDNA sequences of these species and CM04-06 were compared, and then the neighbour-joining phylogenetic tree was drawn by using Mega5 software.

[0148] 1.4 Microbiological characteristics of CM04-06

[0149] (1) Morphological characteristics: CM04-06 was cultured in anaerobic environment at 37°C for 2-3 days, the colony was yellowish white, with high water content, slightly sticky, nearly round, opaque, flat with convex middle, and the colony diameter was about 2-3mm ( Figure 1 )。

[0150] (2) Microscopic characteristics: The cell of CM04-06 was long rod-shaped under 1000 times microscopic magnification, and the Gram staining reaction was negative. No spore and flagellum was found. The diameter of the cell was about 1 um, and the length was 4-10 um. Figure 2 ).

[0151] (3) Physiological and biochemical characteristics: The oxidase and catalase reactions were both negative. The growth temperature range was 30-45 °C, and the pH value range was 4.0-9.0. The optimum temperature and pH value were 37 °C and pH 7.0, respectively. CM04-06 could tolerate 3% NaCl. The comparison of physiological and biochemical reactions (including substrate utilization API 20A and enzyme reaction API ZYM) between CM04-06 and the closest relative reference strain ATCC 27768 was as follows: (+, positive reaction; -, negative reaction; w, weak positive reaction).

[0152] Table 2

[0153]

[0154]

[0155] The comparison of physiological and biochemical reactions between CM04-06 and the reference strains showed that CM04-06 was significantly different from the reference strains in the utilization of raffinose and trehalose, the hydrolysis of aescin and gelatin, and the activities of esterase (C4), valine arylamidase, cystine arylamidase, chymotrypsin, naphthol-AS-BI-phosphohydrolase, β-glucuronidase, a-glucosidase, and N-acetyl- glucosaminidase. It could be proved that CM04-06 was a new species different from the known bacteria.

[0156] 1.5 Analysis of cell fatty acids

[0157] The cells of CM04-06 and Faecalibacterium prausnitzii ATCC 27768 (hereinafter referred to as ATCC 27768, purchased from American Type Culture Collection, and the preservation number was ATCC 27768) cultured to the stationary phase were collected, and then the extraction and detection of cell fatty acids were performed. The contents and differences of cell fatty acid components of the two strains were analyzed by gas chromatography.

[0158] Table 3

[0159]

[0160]

[0161] Based on the above CM04-06 phenotypes, 16S rDNA sequence and physiological and biochemical reactions, CM04-06 is a new species, which is named Faecalibacterium longum.

[0162] Example 2 Bioactive substance of Faecalibacterium longum CM04-06

[0163] 2.1 Detection of short chain fatty acid (SCFA)

[0164] (1) Sample preparation: 1ml of CM04-06 bacterial solution cultured for 48h was centrifuged at 12000r / min for 5min, and the supernatant was taken for standby.

[0165] (2) Determination of SCFA: The determination of short chain fatty acid used external standard method, and acetic acid, propionic acid, butyric acid and valeric acid were selected to make standard curve. Agilent gas chromatograph (GC-7890B, Agilent) was used, and HP-INNOWax (Cross-Linked PEG) 30m x 0.25mm x 0.25um capillary column was selected for analysis, and hydrogen flame ionization detector was used, and GC parameters were set as follows: column temperature: 180-200℃; gasification chamber temperature: 240℃; detection temperature: 210℃; sample amount: 2μL; carrier gas flow: N2, 50mL / min; hydrogen flow: 50mL / min; air flow: 600-700mL / min.

[0166] (3) Results: The SCFA production was determined as follows: formic acid (7.62mmol / L), acetic acid (44.8mmol / L), and butyric acid (40.03mmol / L).

[0167] 2.2 Detection of organic acid

[0168] (1) Sample preparation: same as part 2.1.

[0169] (2) Determination of organic acid: the detection standard of organic acid was selected as 3-methylbutyric acid, valeric acid, quinic acid, lactic acid, oxalic acid, malonic acid, benzoic acid, maleic acid, succinic acid, trans-fumaric acid, malic acid, adipic acid, tartaric acid, shikimic acid, citric acid, isocitric acid and L-ascorbic acid. Agilent gas chromatograph (GC-7890B, Agilent) was still used, and 122-5532G DB-5ms (40m x 0.25mm x 0.25um) was selected as the chromatographic column, and the column temperature was 270-290℃; the sample inlet temperature was 250℃; and the gas flow was 0.86mL / min.

[0170] (3) Results: The organic acid production was determined as shown in the following table (Table 4).

[0171] Table 4

[0172] Organic acid 3-methylbutyric acid Valeric acid Quinic acid Lactic acid Oxalic acid Malonic acid Content (mmol / L) 0.26 0.49 0 30.53 0 0 Organic acid Benzoic acid Maleic acid Succinic acid Trans-fumaric acid Malic acid Adipic acid Content (mmol / L) 1.79 0 0.88 0 0 0.60 Organic acid Tartaric acid Shikimic acid Citric acid Iso-citric acid L-ascorbic acid Content (mmol / L) 0 0 0. 0 0.11

[0173] Example 3, Detection of Exopolysaccharide Produced by Faecalibacterium longum CM04-06

[0174] The detection of exopolysaccharide uses the sulfuric acid phenol method, which can produce a color reaction with free monosaccharides, oligosaccharides and polysaccharides such as hexoses, and the color produced is directly proportional to the absorbance, with an absorption wavelength of 490 nm. The specific experimental process is as follows:

[0175] (1) Extraction of polysaccharides

[0176] The experimental strain was cultured in PYG medium (formula same as Example 1) for 2 days, 10 ml of bacterial solution was treated in a boiling water bath for 30 min, then centrifuged at 10000 r / min, the supernatant was taken, 80% trichloroacetic acid was added to a final concentration of 8%, and it was treated overnight at 4°C to precipitate the protein. The pH of the supernatant was adjusted to 6.0 with NaOH. Add 2 times the volume of anhydrous ethanol for polysaccharide precipitation, treat overnight at 4°C, take out and centrifuge at 10000 r / min for 30 min, discard the supernatant, and dissolve the precipitate with preheated distilled water, then transfer to an ultrafiltration tube (3000 Da filter diameter) for ultrafiltration, centrifuge at 5000 r / min for 30 min, transfer the polysaccharide trapped in the inner tube to a volumetric flask and dilute to 10 ml with distilled water, and reserve.

[0177] (2) Preparation of glucose standard curve

[0178] Precisely weigh 20 mg of standard glucose into a 100 ml volumetric flask, add water to the mark, then prepare 20, 40, 60, 80, 100 μg / ml glucose standard solutions. Take 400 ul of each group of standard solution, three parallel, add 400 ul of 5% phenol and 1 ml of concentrated sulfuric acid in turn for reaction, after boiling water bath for 15 min, cool to room temperature, measure the absorbance at 490 nm. Then take the absorbance as the ordinate, and the glucose standard solution concentration as the abscissa to draw the standard curve.

[0179] (3) Detection of the concentration of extracted polysaccharides

[0180] Take 400 ul of polysaccharide solution, add 400 ul of 5% phenol and 1 ml of concentrated sulfuric acid in turn for reaction, after boiling water bath for 15 min, cool to room temperature, measure the absorbance at 490 nm. Calculate the concentration of polysaccharides according to the glucose standard curve.

[0181] (4) Results

[0182] By calculation, the content of exopolysaccharide in the fermentation broth of CM04-06 cultured for 2 days was 233 mg / L.

[0183] Example 4 In vivo test of Faecalibacterium longum CM04-06 in a diabetic animal model

[0184] This example uses high-fat feed and injection of streptozotocin (STZ) to induce a type 2 diabetes (T2D) model mouse, and investigates the therapeutic effect of CM04-06 on type 2 diabetes (T2D) by gavage with Faecalibacterium longum CM04-06 for 2 months.

[0185] 4.1 T2D mouse modeling: The test mice were C57bL / 6 mice (purchased from Hubei Medical Experimental Animal Center), 8 weeks old, and the mouse test environment was SPF level. The mice were adaptively fed for 1 week and grouped for modeling. The modeling method used high-fat plus STZ induction, and a fasting blood glucose value (FBG) of 10 mM / L or higher was used as a T2D model mouse.

[0186] 4.2 Experimental grouping:

[0187] (1) Model (Model) group: T2D model mice were gavaged with normal saline;

[0188] (2) CM04-06 treatment group: T2D model mice were treated by gavage with CM04-06 bacterial solution;

[0189] (3) Positive drug group: T2D model mice were treated with the positive drug metformin.

[0190] 4.3 Preparation of bacterial solution: CM04-06 was cultured to the stationary phase, to a bacterial concentration of about 10 8 cfu / ml order of magnitude.

[0191] 4.4 Test process: The experiment was carried out according to the grouping, and treatment was started after the T2D model was established (FBG > 10) and lasted for 2 months. The mice's eating activity was recorded daily, and the body weight was measured. Tail vein blood was collected every week to detect the fasting blood glucose value of the mice. The mice's glucose tolerance (OGTT) was measured at the fourth week of treatment and at the end of the experiment. After 2 months of testing, the mice were sacrificed, and the blood was collected from the eyeball, the neck was removed, and the serum was detected for total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C).

[0192] 4.5 Experimental results and analysis

[0193] (1) Body weight change: As shown in Tables 5 and Figure 3As shown, the body weight of the mice gradually increased as the experiment progressed, and the increase in body weight of the mice in the CM04-06 and metformin treatment groups was lower than that in the model group, thus indicating that CM04-06 can effectively slow the increase in body weight of the mice (*P<0.05 / **P<0.01). Meanwhile, statistics show that CM04-06 is more effective than metformin in controlling the increase in body weight of the mice.

[0194] Table 5

[0195] Group number 1 week 3 weeks 5 weeks 7 weeks 9 weeks Model group (g) 22.48 24.99 28.42 31.81 33.45 CM04-06 group (g) 21.56 22.76 24.59 25.41 25.65** Metformin group (g) 21.54 23.42 25.23 26.6 27.49*

[0196] (2) Experimental results of CM04-06 on fasting blood glucose of mice

[0197] Table 6

[0198] Group number First week Third week Fifth week Seventh week Ninth week Model group (mmol / L) 15.5 16.8 17.2 16.9 15.2 CM04-06 group (mmol / L) 16.2 15.2 13.4* 10.8** 8.2** Metformin group (mmol / L) 16.7 15.8 14.6 12.1* 9.8**

[0199] According to Table 6 and Figure 4 The results show that the fasting blood glucose values of the mice in the CM04-06 and metformin groups gradually decreased as the treatment progressed, and the blood glucose values of the mice tended to be normal, and the hypoglycemic effect was relatively significant, indicating that CM04-06 can effectively reduce blood glucose (compared with the model group, *P value <0.05 / **P<0.01). Meanwhile, the blood glucose values of the mice in the CM04-06 group were lower than those in the metformin group, and it can be seen that the ability of CM04-06 to reduce fasting blood glucose is superior to that of metformin.

[0200] (3) Experimental results of CM04-06 on glucose tolerance of mice

[0201] Through OGTT testing of the mice before sacrifice (Table 7 and Figure 5 ), the results show that after intragastric administration of glucose, the blood glucose value of the mice reached the highest (18.2-22.5 mmol / L) at 30 min, and then the blood glucose values of the mice in the metformin and CM04-06 groups decreased at a uniform rate, reaching 9.8 mmol / L and 10.1 mmol / L at 120 min, while the blood glucose value of the mice in the model group was 15.2. At this time point, the blood glucose value of the mice in the CM04-06 group was significantly different from that in the model group (*P value <0.05), and in comparison, CM04-06 can effectively improve the glucose intolerance of diabetic mice. In the whole process of glucose regulation, the blood glucose values of the mice in the CM04-06 group at each time point were lower than those in the metformin group, and thus it can be seen that the improvement effect of CM04-06 on the glucose tolerance of diabetic mice is better.

[0202] Table 7

[0203] Group number 0 min 30 min 60 min 90 min 120 min Model group (mmol / L) 15.8 22.5 17.2 15.8 15.2 CM04-06 group (mmol / L) 9 18.2 15 10.8* 9.8* Metformin group (mmol / L) 9.2 19.5 16.1 10.9* 10.1*

[0204] (4) Effect of CM04-06 on blood lipids of mice

[0205] By detecting the blood lipids of mice after the experiment, including total cholesterol (TC), triglyceride (TG), low density lipoprotein (LDLC) and high density lipoprotein (HDLC), the detection results are shown in Table 8. The results show that the low density lipoprotein of the metformin group and the CM04-06 group is lower than that of the model group (*P value <0.05), and at the same time, the high density lipoprotein is higher than that of the model group (*P value <0.05). The main function of high density lipoprotein is to remove excessive cholesterol and low density lipoprotein in blood and cells, and has the effect of anti-atherosclerosis. Therefore, it can be seen that CM04-06 can improve the hyperlipidemia symptoms of diabetic mice. At the same time, compared with the metformin group, the improvement effect of CM04-06 on blood lipids is better, including lower content of low density lipoprotein and higher content of high density lipoprotein.

[0206] Table 8

[0207]

[0208] Example 5 Food composition containing Faecalibacterium longum CM04-06

[0209] The raw material ratio is shown in Table 9.

[0210] Table 9

[0211]

[0212]

[0213] According to the above formula ratio, mix milk and sugar, stir until completely mixed, preheat, 20Mpa pressure homogenization, sterilize at about 90℃ for 5-10 minutes, cool to 40-43℃, mix in vitamin C, inoculate 1-100×10 6 cfu / g of Faecalibacterium longum CM04-06 bacteria, to make a food composition containing Faecalibacterium longum CM04-06 bacteria.

[0214] Example 6 Pharmaceutical composition containing Faecalibacterium longum CM04-06

[0215] The raw material ratio is shown in Table 10.

[0216] Table 10

[0217]

[0218] Lactose, yeast powder, peptone were mixed with purified water in proportion, preheated to 60-65℃, homogenized under 20Mpa pressure, sterilized at about 90℃ for 20-30 minutes, cooled to 36-38℃, mixed with vitamin C, inoculated with Faecalibacterium longum CM04-06 live bacteria (1-50×10 6 cfu / mL), fermented at 36-38℃ until the pH value was 6.0, centrifuged, freeze-dried to less than 3% moisture content, to prepare Faecalibacterium longum CM04-06 freeze-dried material. 0.5g of Faecalibacterium longum CM04-06 freeze-dried material was mixed with an equal amount of malt dextrin and then loaded into a capsule to prepare a pharmaceutical composition containing Faecalibacterium longum CM04-06 bacteria.

[0219] Example 7 Preparation of a drug containing Faecalibacterium longum CM04-06

[0220] 7.1 Preparation of bacterial solution: Faecalibacterium longum CM04-06 (1×10 9 cfu / ml) was anaerobically cultured, and the anaerobic culture medium was PYG medium, which was fermented anaerobically at 37℃ for 2-3 days.

[0221] 7.2 Preparation of growth factors: skimmed milk and casein were mixed, centrifuged, and ultrafiltered to obtain crude milk growth factor extract (nutrients containing vitamin substances, purine substances, and / or pyrimidine substances).

[0222] 7.3 Preparation of drug dosage form: 5 volumes of the above growth factors were added to 100 volumes of CM04-06 fermented bacterial solution, thoroughly stirred and mixed, and then starch excipients (such as malt dextrin) were added, to prepare a drug containing Faecalibacterium longum CM04-06.

[0223] Preservation of strains

[0224] The strain Faecalibacterium longum CM04-06 of the present application (same as the preservation name) was preserved in the China General Microbiological Culture Collection Center (CGMCC) on June 13, 2016, at No. 1, Beichen West Road, Haidian District, Beijing, China, with the preservation number: CGMCC 1.5208.

[0225] All documents referred to in this disclosure are incorporated herein by reference as if each were individually incorporated by reference. In addition, it is to be understood that various alterations and modifications can be made to the disclosure herein, and it is intended that the same be considered as falling within the scope of the application as defined by the appended claims. SEQUENCE LISTING <110> Shenzhen Huada Gene Research Institute <120> Faecalibacterium longum and its application <130> P2018-1409 <150> PCT / CN2016 / 098246 <151> 2016-09-06 <160> 3 <170> PatentIn version 3.5 <210> 1 <211> 1372 <212> DNA <213> Faecalibacterium longum CM04-06 <400> 1 caagtcgaac gagagatgag gagcttgctc ttcagatcga gtggcgaacg ggtgagtaac 60 gcgtgaggaa cctgcctcaa agagggggac aacagttgga aacgactgct aataccgcat 120 aagcccacgg ctcggcatcg agcagaggga aaaggagtga tccgctttga gatggcctcg 180 cgtccgatta gctggttggt gaggtaacgg cccaccaagg cgacgatcgg tagccggact 240 gagaggttga acggccacat tgggactgag acacggccca gactcctacg ggaggcagca 300 gtggggaata ttgcacaatg ggggaaaccc tgatgcagcg acgccgcgtg gaggaagaag 360 gtcttcggat tgtaaactcc tgttgttgag gaagataatg acggtactca acaaggaagt 420 gacggctaac tacgtgccag cagccgcggt aaaacgtagg tcacaagcgt tgtccggaat 480 tactgggtgt aaagggagcg caggcgggag aacaagttgg aagtgaaatc catgggctca 540 acccatgaac tgctttcaaa actgtttttc ttgagtagtg cagaggtagg cggaattccc 600 ggtgtagcgg tggaatgcgt agatatcggg aggaacacca gtggcgaagg cggcctactg 660 ggcaccaact gacgctgagg ctcgaaagtg tgggtagcaa acaggattag ataccctggt 720 agtccacacc gtaaacgatg attactaggt gttggaggat tgaccccttc agtgccgcag 780 ttaacacaat aagtaatcca cctggggagt acgaccgcaa ggttgaaact caaaggaatt 840 gacgggggcc cgcacaagca gtggagtatg tggtttaatt cgacgcaacg cgaagaacct 900 taccaagtct tgacatccct tgacgaacat agaaatattt tttctcttcg gagcaaggag 960 acaggtggtg catggttgtc gtcagctcgt gtcgtgagat gttgggttaa gtcccgcaac 1020 gagcgcaacc cttatggtca gttactacgc aagaggactc tggccagact gccgttgaca 1080 aaacggagga aggtggggat gacgtcaaat catcatgccc tttatgactt gggctacaca 1140 cgtactacaa tggcgttaaa caaagagaag caagaccgcg aggtggagca aaactcagaa 1200 acaacgtccc agttcggact gcaggctgca actcgcctgc acgaagtcgg aattgctagt 1260 aatcgtggat cagcatgcca cggtgaatac gttcccgggc cttgtacaca ccgcccgtca 1320 caccatgaga gccgggggga cccgaagtcg gtagtctaac cgcaaggagg ac 1372 <210> 2 <211> 20 <212> DNA <213> artificial sequence <400> 2 agagtttgat catggctcag 20 <210> 3 <211> 22 <212> DNA <213> artificial sequence <400> 3 tagggttacc ttgttacgac tt 22

Claims

1. Faecalibacterium prausnitzii, characterized in that, The Faecalibacterium longum is Faecalibacterium longum CM04-06, and the preservation number is CGMCC NO. 1.5208.

2. Faecalibacterium prausnitzii according to claim 1, characterized in that, The sequence of the 16s rDNA of the Faecalibacterium longum is shown as SEQ ID NO:

1.

3. A composition characterized in that, The composition comprises: (a) a safe and effective amount of the Faecalibacterium longum of claim 1; and (b) a food acceptable carrier or a pharmaceutically acceptable carrier.

4. The composition of claim 3, wherein The composition contains 1 x 10 -1 x 10 10 cfu / mL or cfu / g Faecalibacterium longum CM04-06, per total volume or total weight of the composition.

5. The composition of claim 3, wherein The composition further comprises other probiotics and / or prebiotics.

6. Use of Faecalibacterium prausnitzii or a composition containing Faecalibacterium prausnitzii for the manufacture of a formulation for one or more uses selected from the group consisting of: (a) preventing and / or treating obesity; (b) reducing blood lipids; and / or (c) preventing and / or treating diabetes, wherein, The Faecalibacterium longum is Faecalibacterium longum, and the Faecalibacterium longum is the Faecalibacterium longum of claim 1 or 2.

7. Use according to claim 6, characterized in that, The preparation comprises a microecological preparation.

8. Use of the Faecalibacterium longum or the composition containing the Faecalibacterium longum in the preparation of a preparation for one or more purposes selected from the group consisting of: (i) inhibiting weight gain of a mammal; (ii) reducing blood lipid level of a mammal; (iii) increasing the level of high-density lipoprotein (HDL-C) in a mammal; (iv) reducing the level of low-density lipoprotein (LDL-C) in a mammal; (v) reducing blood glucose level of a mammal; (vi) improving sugar tolerance of a mammal; wherein The Faecalibacterium longum is Faecalibacterium longum, and the Faecalibacterium longum is the Faecalibacterium longum of claim 1 or 2.

9. The use of claim 8, wherein, The reduction of blood lipid level of a mammal comprises reducing the level of total cholesterol (TC) and / or the level of triglyceride.

10. A process for the preparation of the composition of claim 3, characterized in that, The method comprises the steps of: mixing the Faecalibacterium longum of claim 1 with a food acceptable carrier or a pharmaceutically acceptable carrier, thereby forming the composition of claim 3.

11. The manufacturing method as described in claim 10, characterized in that, The method further comprises the step of mixing with a growth factor.

12. A production method characterized by comprising: The method comprises the steps of: (a) culturing the Faecalibacterium longum of claim 1 under conditions suitable for growth, thereby obtaining a culture product; (b) isolating the Faecalibacterium longum cells from the culture product; and (c) mixing the Faecalibacterium longum cells isolated in the previous step with a food acceptable carrier or a pharmaceutically acceptable carrier, thereby preparing the composition.

13. The production method according to claim 12, wherein Before step (c), the method further comprises the step of mixing the isolated Faecalibacterium longum cells with a growth factor.

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