CN115074277A

CN115074277A - Clostridium pralatanorum capable of relieving obesity and application thereof

Info

Publication number: CN115074277A
Application number: CN202210728189.0A
Authority: CN
Inventors: 张灏; 翟齐啸; 代步珅
Original assignee: Wuxi Special Food And Nutrition Health Research Institute Co ltd
Current assignee: Wuxi Special Food And Nutrition Health Research Institute Co ltd
Priority date: 2022-06-24
Filing date: 2022-06-24
Publication date: 2022-09-20
Anticipated expiration: 2042-06-24
Also published as: CN115074277B

Abstract

The invention relates to clostridium pralatanorum capable of relieving obesity and application thereof, and belongs to the technical field of microorganisms and medicines. The clostridium prausnitzii (Faecalibacterium praussnitzii) CCFM1203 of the invention has the function of relieving obesity, and is specifically reflected in that: (1) significantly inhibiting weight gain; (2) remarkably reducing the weight of abdominal fat and inhibiting the expansion of fat cells; (3) the content of TC, TG, FFA and LDL-C in serum of an obese mouse is remarkably reduced, and the content of HDL-C is improved, so that the Clostridium prausnitzii (Faecalibacterium prausnitzii) CCFM1203 has a huge application prospect in preparation of products for preventing and/or treating obesity.

Description

Clostridium pralatanorum capable of relieving obesity and application thereof

Technical Field

The invention relates to clostridium pralatanorum capable of relieving obesity and application thereof, and belongs to the technical field of microorganisms and medicines.

Background

Obesity is a chronic disease and seriously harms human health. Obesity can increase the risk of a variety of diseases, including cardiovascular disease, diabetes, musculoskeletal disorders, and cancer, among others. The World Health Organization (WHO) defines obesity as an abnormal or excessive fat accumulation that can impair health, and generally considers BMI indices in excess of 25kg/m ² Is considered overweight, more than 30kg/m ² It is obese. Currently, about 13% of adults worldwide (11% in men, 15% in women) have the characteristic of being obese. The prevalence of obesity has increased nearly three times worldwide since 1975. The lack of physical activity due to excessive intake of high calorie food rich in fat and sugar by modern people, and the increase in urbanization and the change in transportation means are all causes of the increase in global obesity rate.

Obesity and the underlying cause are an imbalance between intake of calories and energy before consumption of calories, usually an excess of body fat due to changes in its physiological or biochemical functions. From the pathogenesis, the obesity has two types; 1) simple obesity; 2) secondary obesity. Simple obesity can be divided into congenital obesity and acquired obesity, and the number of patients with simple obesity can be more than 95% of the total obesity. Congenital obesity is caused by a large number of fat cells and is common in children; then, the acquired obesity is caused by the massive expansion of fat cells and is commonly seen in adults. Secondary obesity, also known as symptomatic obesity, is usually caused by endocrine or metabolic disorders.

Recent advances in basic and applied knowledge of the human gut microbiota have changed the current opinion on obesity and promising treatment modalities. The human intestinal microbiota has been identified as a complex ecosystem comprising a number of bacterial species, the number of which exceeds 1000Trillion (10) ¹⁴ Order of magnitude) of 10 times or more the total number of human cells. In the long evolution process, the intestinal microorganisms achieve good mutual benefit relationship with human beings, and play an important role in nutrition, metabolism and immunity of the human bodies. Studies have shown that the intestinal flora of obese patients is reduced in beneficial bacteria and increased in harmful bacteria compared to the intestinal flora of healthy people, and that the metabolites in the intestine are altered, such as reduced short chain fatty acids.

Currently, the main strategies for treating obesity are: diet regulation, physical exercise, behavior therapy, drug therapy, and rehabilitation surgery. The means employed will depend primarily on the health risk factors of the patient and the rate and effect of weight loss, and combinations may be selected for the treatment of obese patients. Dietary exercise is a general therapy, namely eating low-calorie and low-fat food combined with aerobic exercise, but the method is generally considered to be unsuccessful for the general public and needs to be frequently maintained for a long time; the operation of removing the fat in the body can play a role in the instant, but has the defects of certain operation risk, difficult fat removal effect and over expensive cost. Medications are commonly taken in the form of phenylpropanolamine (PPA), freshmus rosenbergii (orlistat III) and sibutramine (reduct), the mechanisms of which mainly include appetite suppression, increased energy expenditure, reduced triglycerides and fat absorption inhibition. However, medication often causes certain side effects (e.g., abdominal pain, diarrhea, liver injury, etc.) and tends to rebound once taken.

In response to the disadvantages of the above-mentioned drugs, researchers have begun to attempt to treat obesity using probiotic formulations. The probiotic preparation is safe and healthy to human bodies, and the administration mode is simple and convenient. Representative probiotics include lactobacillus and bifidobacterium, but the probiotics such as lactobacillus and bifidobacterium which are found to relieve obesity at present have poor or undefined effects, while most of the probiotics in the market have the defect of slow effect or undefined action relationship.

Therefore, a probiotic with strong obesity pertinence and strong obesity relieving capability is urgently needed to be found so as to solve the problems of slow effect and unclear action relationship of the existing probiotic preparation for obesity.

Disclosure of Invention

The invention provides a strain of clostridium prausnitzii (clostridium prausnitzii) CCFM1203, wherein the clostridium prausnitzii (clostridium prausnitzii) CCFM1203 is preserved in Guangdong province microorganism culture collection center, and the preservation number is GDMCC No: 62238, the preservation date is 2022, 1 month, 27 days, and the preservation address is No. 59 building 5 of Dazhou college No. 100 of Jieli Zhonglu, Guangzhou city.

In one embodiment, the C.praerusni (Clostridium prausnitzii) CCFM1203 is derived from healthy adult feces, the 16S rDNA sequence of the strain is shown in SEQ ID NO.1 by analysis, and the colony of the strain on the M2GSC medium is circular convex, smooth in surface and translucent.

The invention also provides a product containing the clostridium prausnitzii (clostridium prausnitzii) CCFM 1203.

In one embodiment, the product is a probiotic formulation or a medicament.

In one embodiment, the probiotic formulation contains, in addition to clostridium pralatanolyticum CCFM1203, excipients including, but not limited to, excipients or food additives; the content of the clostridium praeparatum CCFM1203 in the probiotic preparation is not less than 1 x 10 ⁶ CFU/mL or 1X 10 ⁶ CFU/g。

In one embodiment, the medicament is for preventing, ameliorating, improving and/or treating obesity.

In one embodiment, the medicament further comprises a pharmaceutical excipient.

In one embodiment, the medicament is for use in at least one of (a) - (d):

(a) inhibiting weight gain;

(b) reducing the weight of abdominal fat and inhibiting expansion of adipocytes;

(c) improving blood lipid environment, and reducing TC, TG and FFA content in serum;

(d) increasing the transport level of TG in the liver, up-regulating HDL-C levels, and down-regulating LDL-C levels;

in one embodimentThe content of the clostridium prasukii CCFM1203 in the medicine is not less than 1 x 10 ⁶ CFU/mL or 1X 10 ⁶ CFU/g。

In one embodiment, the improvement in the lipid profile comprises down-regulation of TC, TG, LDL-C and FFA levels in the blood, or up-regulation of HDL-C levels.

In one embodiment, the pharmaceutical excipient comprises an excipient and an additive.

In one embodiment, the pharmaceutical excipient comprises an anti-adhesive, an osmotic enhancer, a buffering agent, a plasticizer, a surfactant, an antifoaming agent, a thickener, an encapsulating agent, an absorbent, a humectant, a solvent, a propellant, a solubilizer, a cosolvent, an emulsifier, a colorant, a pH adjuster, a binder, a disintegrant, a filler, a lubricant, a wetting agent, an integrating agent, an osmotic pressure adjuster, a stabilizer, a glidant, a flavoring agent, a preservative, a foaming agent, a suspending agent, a coating material, a fragrance, a diluent, a flocculating agent and a deflocculating agent, a filter aid, and a release retardant.

In one embodiment, the additive comprises microcrystalline cellulose, hydroxypropyl methylcellulose, and refined lecithin.

In one embodiment, the dosage form of the medicament comprises granules, capsules, tablets, pills or oral liquid.

The invention also provides a food, a drink, a health product, an enteral nutrient, a dietary supplement, a veterinary drug or a feed additive containing the clostridium prausnitzii (clostridium prausnitzii) CCFM 1203.

In one embodiment, the food product comprises a dairy, soy, or fruit and vegetable product produced using a starter culture comprising clostridium pralatanolyticum CCFM 1203; or the food product comprises a solid beverage comprising the clostridium pralatanolyticum CCFM 1203.

In one embodiment, the food, nutraceutical, enteral nutrition, dietary supplement, veterinary drug or feed additive further comprises conventional adjuvants.

The invention also provides application of the clostridium pralatanorum CCFM1203 in preparation of food, beverages, health products, enteral nutrition preparations, dietary supplements, veterinary drugs or feed additives.

Has the advantages that:

animal experiments prove that the clostridium prausnitzii (Faecalibacterium prausnitzii) CCFM1203 has the function of relieving obesity, and is specifically shown in the following steps:

(1) the weight of the obese mouse is reduced from 37.50 +/-3.65 g to 30.38 +/-2.28 g;

(2) the abdominal fat weight of the obese mouse is reduced from 1.97 plus or minus 0.55g to 0.81 plus or minus 0.24 g;

(3) the fat cell area of the abdominal cavity of the obese mouse is 8214.77 +/-1357.26 mu m ² The particle size is reduced to 4220.16 +/-681.35 mu m ² ；

(4) The content of the serum HDL-C of the obese mouse is increased from 0.81 plus or minus 0.16mmol/L to 1.33 plus or minus 0.20 mmol/L;

(5) the content of LDL-C in serum of the obese mouse is reduced to 0.50 plus or minus 0.08mmol/L from 0.88 plus or minus 0.17 mmol/L;

(6) the TC content of the serum of the obese mouse is reduced to 3.38 plus or minus 0.51mmol/L from 5.38 plus or minus 0.56 mmol/L;

(7) the serum TG content of the obese mouse is reduced from 1.85 plus or minus 0.28mmol/L to 1.04 plus or minus 0.15 mmol/L;

(8) the FFA content of the serum of the obese mouse is reduced to 1.11 plus or minus 0.21mmol/L from 2.02 plus or minus 0.22 mmol/L;

therefore, the clostridium prausnitzii (clostridium prausnitzii) CCFM1203 has a huge application prospect in preparing products (such as food, medicines or health products and the like) for preventing and/or treating obesity.

Biological material preservation

Clostridium praerussitum (Faecalibacterium praerucitzi) CCFM1203, taxonomically named Faecalibacterium praerucitzi, has been deposited at the guangdong provincial collection of microorganisms at 27 months 1/2022 with the deposit number GDMCC No: 62238, the preservation address is No. 59 building 5 of No. 100 Dazhong Jie-Lu-100 Guangzhou city.

Drawings

FIG. 1: results of body weight of mice tested in different groups.

FIG. 2: results of abdominal fat weight in mice tested in different groups.

FIG. 3: and (3) performing HE staining on abdominal fat of mice in different groups.

FIG. 4 is a schematic view of: results of average areas of adipocytes in abdominal cavities of experimental mice of different groups.

FIG. 5: the content of HDL-C in serum of mice was tested in different groups.

FIG. 6: the content of LDL-C in serum of different groups of experimental mice.

FIG. 7: the content of TC in serum of experimental mice of different groups.

FIG. 8 is the following: the content of TG in serum of different groups of experimental mice.

FIG. 9: the FFA content in the serum of the experimental mice of different groups.

Detailed Description

The invention is further illustrated with reference to the following specific examples.

Skim milk, referred to in the following examples, was purchased from Guangming milk industries, Inc., glucose and yeast extract were purchased from national pharmaceutical group Chemicals, Inc., casein peptone was purchased from Shanghai Chungsai, TC, FFA, TG, HDL-C, and LDL-C kits were purchased from Beckman coulter, USA.

The model strain Clostridium prausnitzii (Faecalibacterium prausnitzii) A2-165 was obtained from German Collection of microorganisms and cell cultures (DSMZ) with the accession number DSMZ 17677.

The media involved in the following examples are as follows:

m2GSC solid medium (g/L): 5g/L of yeast powder, 10g/L of casein peptone, 5g/L of glucose, 2g/L of cellobiose, 2g/L of fructose, 4g/L of sodium bicarbonate, 0.9g/L of sodium chloride, 0.45g/L of potassium dihydrogen phosphate, 0.45g/L of dipotassium hydrogen phosphate, 0.09g/L of magnesium sulfate, 0.09g/L of calcium chloride, 15g/L of agar, 0.5g/L of cysteine, 1.0mg/L of resazurin and 10mL/L of clarified rumen fluid.

M2GSC liquid medium (g/L): 5g/L of yeast powder, 10g/L of casein peptone, 5g/L of glucose, 2g/L of cellobiose, 2g/L of fructose, 4g/L of sodium bicarbonate, 0.9g/L of sodium chloride, 0.45g/L of potassium dihydrogen phosphate, 0.45g/L of dipotassium hydrogen phosphate, 0.09g/L of magnesium sulfate, 0.09g/L of calcium chloride, 0.5g/L of cysteine, 1.0mg/L of resazurin and 10mL/L of clarified rumen fluid.

Example 1: screening and strain identification of clostridium pralatanorum

(1) Screening

Taking healthy human excrement from Wuxi region in Jiangsu as a sample, and immediately transferring the sample into an anaerobic workstation for treatment within one hour after the sample is collected. Taking a spoon of feces sample, adding into 5mL PBS (adding 0.05% cysteine), mixing well, performing gradient dilution, selecting 10 ^-5 ～10 ^-7 The gradient diluent is coated on the M2GSC solid culture medium, cultured for 48h at 37 ℃, a typical colony is picked into two M2GSC solid culture mediums, one M2GSC solid culture medium is placed outside an anaerobic workstation to be cultured (named as an A plate), the other M2GSC solid culture medium is placed outside the anaerobic workstation to be exposed for 30min and then taken back to the anaerobic workstation to be cultured (named as a B plate), the culture temperature is 37 ℃, after being cultured for 48h, a colony which does not grow in the B plate is picked to be streaked on the M2GSC solid culture medium for purification, and specific primer detection is carried out at the same time. After purification, selecting a single colony, transferring the single colony into an M2GSC liquid culture medium, culturing at 37 ℃, performing suction filtration in an anaerobic workstation to collect bacterial sludge, and preserving by 30% glycerol to obtain a strain CCFM 1203.

(2) Identification

Single colonies of CCFM1203 were picked for colony PCR to amplify and sequence their 16s rDNA (by jingzhi). The 16s rDNA sequence of CCFM1203 is SEQ ID NO.1, and the sequence is subjected to nucleic acid sequence alignment in NCBI, so that the strain is clostridium praerussitum and is named as clostridium prausnitzii (Faecalibacterium prausnitzii) CCFM 1203.

Example 2: culture of Clostridium pralatum

After the clostridium praerusinitzii (Faecalibacterium prausnitzii) CCFM1203 is inoculated into the M2GSC solid medium and cultured at 37 ℃ for 48h, the colony is observed, and the colony is convex, translucent, complete in edge and moist.

Inoculating 2% of strain of Clostridium prausnitzii (Clostridium prausnitzii) CCFM1203 into M2GSC liquid culture medium, anaerobically culturing at 37 deg.C for 24 hr, transferring into fresh M2GSC liquid culture medium, culturing under the same condition for 24 hr, vacuum filtering in an anaerobic workstation to obtain bacterial sludge, adding 0.1M PBS (pH 7.2 containing 0.05% of cysteine, adding water, stirring, and stirring to obtain a suspensionAmino acid) washing and re-suspending to obtain a bacterial concentration of 1 × 10 ⁹ CFU/mL bacterial suspension, using good sealing freeze-drying bottle plug preservation bacterial suspension, the day use.

Example 3: effect of Clostridium pralatanorum on body weight of obese mice

32 SPF-grade male C57BL/6J mice (8 weeks old, 18-22g) were randomly divided into 4 groups of 8 mice each, blank, model, A2-165 and CCFM 1203. The mice are bred in the experimental animal center of university in south of the Yangtze river, the constant temperature is 21-26 ℃, the humidity is 40-70%, the noise is less than or equal to 60dB, and the animal illumination is 15-20LX (all animal experimental procedures are examined and approved by animal welfare and ethical management committee of the university in south of the Yangtze river).

The experiment took 13 weeks: the mice are adaptively fed for 7 days, and from day 8, the mice in the model group and the mice in the intervention group are fed with high-fat feed, and the mice in the blank group are still fed with common feed. In addition, the mice in the intervention group were gavaged with 0.2mL of Clostridium prasukii CCFM1203 and Clostridium prasukii model strain A2-165 bacterial solution (1X 10) each day from day 8 ⁹ mL), blank and model groups were gavaged with equal volumes of PBS solution.

The body weight of the mice was measured weekly during the experiment and the results are shown in figure 1.

As shown in fig. 1, the body weight of the model group mice was significantly increased to 37.50 ± 3.65g compared to 26.35 ± 1.22g of the blank group. After the mice are fed with the strain clostridium prausnitzii (Faecalibacterium prausnitzii) CCFM1203, the body weight is obviously reduced by 18.98 percent compared with the model group (P <0.05), and is close to the body weight of the normal group; the mice in the A2-165 group are reduced by 10.99%, which shows that the Clostridium prausnitzii (Faecalibacterium prausnitzii) CCFM1203 can reduce the weight of obese mice, and the weight reducing effect is obviously better than that of the model strain A2-165(P < 0.05).

Example 4: effect of Clostridium pralatanorum on abdominal fat weight in obese mice

After the experiment was completed, blood was taken and the mice were sacrificed, abdominal fat was removed and weighed, and the results are shown in fig. 2.

As shown in FIG. 2, the abdominal fat weight of the model mice was significantly increased to 1.97. + -. 0.55g compared to 0.33. + -. 0.07g of the blank group. After mice were fed clostridium prausnitzii CCFM1203 of the present invention, the fat weight decreased significantly 58.63% compared to the model group (P < 0.05); the mice in the group A2-165 are reduced by 36.84%, and the experiments show that the Clostridium prausnitzii (Faecalibacterium prausnitzii) CCFM1203 can obviously reduce the abdominal fat weight of obese mice, and the effect is obviously better than that of the model strain A2-165(P < 0.05).

Example 5: effect of Clostridium pralatanorum on abdominal fat pathology in obese mice

After the experiment is finished, blood is taken out and a mouse is killed, abdominal fat is taken down and is placed in 4% paraformaldehyde solution for fixation for 36h, after ethanol gradient dehydration, xylene transparence, paraffin embedding and section HE staining, the pathological degree of the tissue is evaluated, the staining result is shown in figure 3, and the fat cell measuring area is shown in figure 4.

As shown in fig. 3, the volume of adipose cells in abdominal adipose tissues of the model group mice was significantly increased compared to that of the blank group; the fat cell volume of the abdominal fat tissue of the mice in the intervention group is reduced to a different extent compared with the model group. As shown in fig. 4, the planar area of the abdominal fat cells of CCFM1203 mice was significantly decreased by 48.63% (P <0.05) compared to the model group; the mice in the group A2-165 are reduced by 35.95%, which shows that the Clostridium prausnitzii (Faecalibacterium prausnitzii) CCFM1203 can inhibit the increase of abdominal fat cells of obese mice, and the effect is obviously better than that of the model strain A2-165(P < 0.05).

Example 6: effect of Clostridium pralatanorum on serum HDL-C of obese mice

32 SPF-grade male C57BL/6J mice (8 weeks old, 18-22g) were randomly divided into 4 groups of 8 mice each, blank, model, A2-165 and CCFM 1203. The mice are bred in experimental animal centers of university in south of the Yangtze river, the constant temperature is 21-26 ℃, the humidity is 40-70%, the noise is less than or equal to 60dB, and the animal illumination is 15-20LX (all animal experimental procedures are reviewed and approved by animal welfare and ethical administration committee of the university in south of the Yangtze river).

After the experiment is finished, blood is taken and mice are killed, the blood is placed at room temperature for 1h, then is centrifuged at 3500r/min at 4 ℃ for 10min, serum is collected, and the content of HDL-C in the serum is measured by a kit, and the result is shown in figure 5.

As shown in FIG. 5, the HDL-C content in the model group mice was significantly reduced to 0.81. + -. 0.16mmol/L (P <0.05) compared to 1.65. + -. 0.27mmol/L in the blank group. After mice are fed with the strain clostridium prausnitzii (Faecalibacterium prausnitzii) CCFM1203 of the invention, the HDL-C content is significantly increased by 63.62% compared with the model group (P < 0.05); the mice in the group A2-165 are improved by 35.94%, which shows that the Clostridium prausnitzii (Faecalibacterium prausnitzii) CCFM1203 can improve the HDL-C content of obese mice, and the effect is obviously better than that of the model strain A2-165(P < 0.05).

Example 7: effect of Clostridium pralatanorum on serum LDL-C in obese mice

The method comprises the following specific steps:

After the experiment, blood is taken and mice are killed, the blood is placed at room temperature for 1h, then is centrifuged at 3500r/min at 4 ℃ for 10min, serum is collected, and the content of LDL-C in the serum is measured by a kit, and the result is shown in figure 6.

As shown in FIG. 6, the LDL-C content in the model group mice was significantly increased to 0.88. + -. 0.17mmol/L (P <0.05) compared to 0.28. + -. 0.04mmol/L in the blank group. After mice were fed with the strain of the invention, clostridium prausnitzii (Faecalibacterium prausnitzii) CCFM1203, the LDL-C content was significantly reduced by 43.28% compared to the model group (P < 0.05); the mice in the A2-165 group are reduced by 24.24%, which shows that the Clostridium prausnitzii (Faecalibacterium prausnitzii) CCFM1203 can reduce the LDL-C content of obese mice, and the effect is obviously better than that of the model strain A2-165(P < 0.05).

Example 8: effect of Clostridium pralatanorum on serum TC of obese mice

The experiment took 13 weeks: the mice were adaptively fed for 7 days, and from day 8, the mice in the model group and the mice in the intervention group were fed with high fat diet, and the mice in the blank group were fed with normal diet. In addition, the mice in the intervention group were gavaged with 0.2mL of Clostridium prasukii CCFM1203 and Clostridium prasukii model strain A2-165 bacterial solution (1X 10) each day from day 8 ⁹ mL), blank and model groups were gavaged with equal volumes of PBS solution.

After the experiment, blood is taken and mice are killed, the blood is placed at room temperature for 1h, then is centrifuged at 3500r/min at 4 ℃ for 10min, serum is collected, and the TC content in the serum is measured by a kit, and the result is shown in figure 7.

As shown in FIG. 7, the TC content of the model group mice significantly increased to 5.38 + -0.56 mmol/L (P <0.05) compared to 1.97 + -0.40 mmol/L of the blank group. After mice were fed with the strain clostridium prausnitzii (Faecalibacterium prausnitzii) CCFM1203 of the present invention, the TC content was significantly reduced by 37.16% compared to the model group (P < 0.05); the mice in the A2-165 group have a 21.93% reduction, which shows that the Clostridium prausnitzii (Faecalibacterium prausnitzii) CCFM1203 can reduce the TC content of obese mice, and the effect is obviously better than that of the model strain A2-165(P < 0.05).

Example 9: effect of Clostridium plazae on serum TG in obese mice

The method comprises the following specific steps:

After the experiment, blood was taken and the mice were sacrificed, the blood was left at room temperature for 1 hour, centrifuged at 3500r/min at 4 ℃ for 10min, serum was collected, and the TG content in the serum was measured by a kit, and the results are shown in fig. 8.

As shown in FIG. 8, TG levels in the model group mice significantly increased to 1.85. + -. 0.28mmol/L (P <0.05) compared to 0.84. + -. 0.13mmol/L in the blank group. After mice are fed with the strains of clostridium prausnitzii (Faecalibacterium prausnitzii) CCFM1203 and A2-165 in the invention, the TG content is reduced by 44.15 percent (P <0.05) and 27.90 percent respectively compared with the model group, which shows that the clostridium prausnitzii CCFM1203 can reduce the TG content of obese mice and the effect is obviously better than that of the clostridium practicum model strain A2-165.

Example 10: effect of Clostridium pralatanorum on serum FFA of obese mice

The experiment took 13 weeks: mice were acclimatized for 7 days, starting on day 8The mice in the model group and the mice in the intervention group were fed with high fat diet, and the mice in the blank group were fed with normal diet. In addition, the mice in the intervention group were gavaged with 0.2mL of Clostridium prasukii CCFM1203 and Clostridium prasukii model strain A2-165 bacterial solution (1X 10) each day from day 8 ⁹ mL), blank and model groups were gavaged with equal volumes of PBS solution.

After the experiment, blood is taken and mice are killed, the blood is placed at room temperature for 1h, then is centrifuged at 3500r/min at 4 ℃ for 10min, serum is collected, and the FFA content in the serum is measured by a kit, and the result is shown in figure 9.

As shown in FIG. 9, FFA content in the model group mice significantly increased to 2.02. + -. 0.22mmol/L (P <0.05) compared to 0.97. + -. 0.20mmol/L in the blank group. After the mice are fed with the strain clostridium prausnitzii (clostridium prausnitzii) CCFM1203 of the invention, the FFA content is obviously reduced by 45.24 percent compared with a model group (P <0.05), which indicates that the clostridium prausnitzii (clostridium prausnitzii) CCFM1203 can reduce the FFA content of obese mice and the effect is obviously better than that of the clostridium practicum model strain A2-165.

Example 11: preparation of solid beverage containing clostridium praecox CCFM1203

The method comprises the following specific steps:

inoculating clostridium pralatum CCFM1203 into a culture medium according to an inoculation amount accounting for 3% of the total mass of the culture medium, and culturing at 37 ℃ for 18 hours to obtain a culture solution; centrifuging the culture solution to obtain thalli; washing the thallus with phosphate buffer solution with pH of 7.2 for 3 times, and re-suspending with trehalose lyophilized protectant with trehalose concentration of 100g/L (mass ratio of lyophilized protectant to thallus is 2:1) to obtain thallus concentration of 5 × 10 ⁸ CFU/mL of the resuspension solution; and (3) freeze-drying the heavy suspension by adopting a vacuum freezing method to obtain the clostridium pralatanorum CCFM1203 bacteria powder.

Will contain 10 ⁹ The powder of the clostridium pralatanorum CCFM1203 in CFU/g is mixed with maltodextrin at a ratio of 2:1 to obtain the solid beverage rich in the clostridium praecox CCFM 1203.

Taking 10g (equivalent to a total irrigation 10) ¹⁰ CFU) the solid beverage containing the clostridium praecox CCFM1203 is dissolved in normal saline again, the volume is fixed to 20 milliliters, each mouse is subjected to intragastric administration of 200 microliters every day for 12 weeks, and the effect of inhibiting the growth of the bacteria can be effectively achievedIncrease body weight and expand fat cells, increase HDL-C content, and reduce TC, TG, FFA and LDL-C content, thereby relieving obesity.

Example 12: preparation of cow milk containing clostridium praecox CCFM1203

The method comprises the following specific steps:

inoculating clostridium pralatanorum CCFM1203 into a culture medium according to the inoculation amount accounting for 3% of the total mass of the culture medium, and culturing at 37 ℃ for 18h to obtain a culture solution; centrifuging the culture solution to obtain thalli; washing the bacteria with phosphate buffer solution with pH of 7.2 for 3 times, and suspending with trehalose lyophilized protectant with trehalose concentration of 100g/L until the bacteria concentration reaches 1 × 10 ¹⁰ CFU/mL to obtain a bacterial suspension; placing the suspension at 37 ℃ for 60min for pre-culture, and freeze-drying by adopting a freeze-drying method to obtain a clostridium prasukii CCFM1203 leavening agent; wherein the culture medium comprises 87.7 percent of water, 10 percent of enzyme hydrolysis skim milk, 0.5 percent of glucose, 1.5 percent of tryptone and 0.3 percent of yeast extract solution by mass of the total culture medium; the pH of the medium was 6.8;

sterilizing skim milk at 95 deg.C for 20min, cooling to 4 deg.C, adding fermentation agent of Clostridium pralatum CCFM1203 to make viable bacteria concentration of Clostridium pralatum CCFM1203 in skim milk reach 1 × 10 ⁹ And (3) storing the milk in a refrigerating way at 4 ℃ in a CFU/mL manner to obtain the milk containing the viable bacteria of the clostridium praecox CCFM 1203.

200 microliters of cow milk containing live bacteria of the clostridium praepanicum CCFM1203 is taken to perform intragastric administration on the mouse for 12 weeks continuously, so that the weight growth and the expansion of fat cells can be effectively inhibited, the content of HDL-C is increased, and the contents of TC, TG, FFA and LDL-C are reduced simultaneously, thereby achieving the effect of relieving obesity.

Although the present invention has been described with reference to the preferred embodiments, it should be understood that various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.

SEQUENCE LISTING

<110> Special tin-free food and Nutrition health research institute Co., Ltd

<120> clostridium pralatanorum capable of relieving obesity and application thereof

<130> BAA2111281A

<160> 1

<170> PatentIn version 3.3

<210> 1

<211> 1378

<212> DNA

<213> Faecalibacterium prausnitzii

<400> 1

gcggcgtcct ccttgcggtt agactaccga cttcgggtcc ccccggctct catggtgtga 60

cgggcggtgt gtacaaggcc cgggaacgta ttcaccgcag catgctgatc tgcgattact 120

agcaattccg acttcgtgca ggcgagttgc agcctgcagt ccgaactggg acgttgtttc 180

tgagttttgc tccacctcgc ggtcttgctt ctctttgttt aacgccattg tagtacgtgt 240

gtagcccaag tcataaaggg catgatgatt tgacgtcatc cccaccttcc tccgttttgt 300

caacggcagt ctggccagag tcctcttgcg tagtaactga ccataagggt tgcgctcgtt 360

gcgggactta acccaacatc tcacgacacg agctgacgac aaccatgcac cacctgtctc 420

tgcgtcccga aggaaacaca tatttctatg tgcgtcgcat gatgtcaaga cttggtaagg 480

ttcttcgcgt tgcgtcgaat taaaccacat actccactgc ttgtgcgggc ccccgtcaat 540

tcctttgagt ttcaaccttg cggtcgtact ccccaggtgg attacttatt gtgttaactg 600

cggcactgaa ggggtcaatc ctccaacacc tagtaatcat cgtttacagt gtggactacc 660

agggtatcta atcctgtttg ctacccacac tttcgagcct cagcgtcagt tggtgcccag 720

taggccgcct tcgccactgg tgttcctccc gatatctacg cattccaccg ctacaccggg 780

aattccgcct acctctgcac tactcaagaa aaacagtttt gaaagcagtt catgggttga 840

gcccatggat ttcacttcca acttgtcttc ccgcctgcgc tccctttaca cccagtaatt 900

ccggacaacg cttgtgacct acgttttacc gcggctgctg gcacgtagtt agccgtcact 960

tccttgttga gtaccgtcat tatcttcctc aacaacagga gtttacaatc cgaagacctt 1020

cttcctccac gcggcgtcgc tgcatcaggg tttcccccat tgtgcaatat tccccactgc 1080

tgcctcccgt aggagtctgg gccgtgtctc agtcccaatg tggccgttca acctctcagt 1140

ccggctaccg atcgtcgcct tggtgggcca ttacctcacc aactagctaa tcggacgcga 1200

ggccatctca aagcggattg ctccttttcc ctctacccga tgccgggtcg tgggcttatg 1260

cggtattagc agtcgtttcc aactgttgtc cccctctttg aggcaggttc ctcacgcgtt 1320

actcacccgt tcgccactcg ctcgagaaag caagctctct ctcgctcgtt cgactgca 1378

Claims

1. A strain of clostridium prausnitzii (clostridium praerusinzii), wherein the clostridium praerucii CCFM1203 is deposited in the collection of microorganisms and strains of Guangdong province at 1 month and 27 months 2022, and the deposit number is GDMCC No: 62238.

2. a microbial preparation comprising said Clostridium prausnitzii (Clostridium prausnitzii) CCFM 1203.

3. The microbial preparation of claim 2, wherein the clostridium pralatum CCFM1203 is present in the microbial preparation in an amount of not less than 1 x 10 ⁶ CFU/mL or 1X 10 ⁶ CFU/g。

4. A medicament containing said clostridium prausnitzii CCFM 1203.

5. The medicament according to claim 4, further comprising an excipient, wherein the excipient includes but is not limited to an excipient or an additive.

6. The medicament of claim 4 or 5, wherein the medicament has at least one of the uses of (a) to (d):

(a) inhibiting weight gain;

(d) increasing the transport level of TG in the liver, up-regulating HDL-C levels, and down-regulating LDL-C levels.

7. Use of clostridium pralatanorum according to claim 1 for the preparation of a medicament for the prevention, alleviation, amelioration and/or treatment of obesity.

8. The use of claim 7, wherein the C.pratensis CCFM1203 is present in the medicament in an amount of not less than 1 x 10 ⁶ CFU/mL or 1X 10 ⁶ CFU/g。

9. Food, beverage, health product, enteral nutrition, dietary supplement, veterinary drug or feed additive containing clostridium praecox CCFM 1203.

10. Use of the clostridium pralatanorum CCFM1203 according to claim 1 in the preparation of a food product, a nutraceutical product, a beverage product, an enteral nutritional preparation, a dietary supplement, a veterinary drug, or a feed additive.