CN118240724B - Bifidobacterium animalis subspecies ProSci-246, products prepared from same and application of products - Google Patents

Abstract

The invention discloses a strain of animal Bifidobacterium lactis subspecies ProSci‑246 and products and applications prepared therefrom, belonging to the technical field of probiotics. The animal Bifidobacterium lactis subspecies ProSci‑246 provided by the present invention is a Gram-positive bacterium, with an optimum growth temperature of 37°C, an optimum growth pH of 6.0, and good tolerance to gastrointestinal fluid; 11 kinds of carbohydrates can be used; no potential toxicity-related genes were detected, and functional genes related to good health were contained; the weight gain and average daily food intake of obese mice can be significantly reduced, the epididymal fat and inguinal fat weight of obese mice are reduced, obese mice tissue inflammation is reduced, and fat cell deposition in the liver and fat is reduced; there is a significant positive effect on the weight, body fat mass, BMI, waist-to-hip ratio, visceral fat index and health score of obese adults, and there is a huge application prospect in products for preventing and/or improving obesity.

Description

A strain of Bifidobacterium animalis subspecies lactis ProSci-246 and products prepared therefrom and their applications

Technical Field

The invention relates to the technical field of probiotics, in particular to a strain of animal Bifidobacterium lactis subspecies ProSci-246 and products prepared therefrom and applications thereof.

Background Art

Probiotics have important physiological functions for the body, such as adjusting the balance of intestinal flora, lowering cholesterol, anti-oxidation, anti-infection and enhancing human immunity, etc. They have broad application prospects in the fields of food and medical health.

As the dominant microorganism in the gastrointestinal tract, bifidobacteria play an irreplaceable role in maintaining intestinal health. In recent years, various clinical, in vivo and in vitro studies have proven that bifidobacteria can play an important role in inflammatory bowel disease, irritable bowel syndrome, cancer, diarrhea, lactose intolerance and other diseases, protecting the integrity of the intestinal barrier, thereby keeping the body healthy and stable.

Obesity is one of the important health problems and is associated with many chronic metabolic diseases, such as hypertension, diabetes and dyslipidemia. According to statistics in 2020, more than 1.9 billion adults in the world face obesity problems. Under the potential influence of environment and genes, the obesity phenomenon has been particularly serious in recent years. Global environmental factors that lead to obesity include reduced exercise, overeating, excessive intake of high-calorie foods, consumption of pickled foods and the use of certain drugs. In addition, obesity is also related to some urological diseases, such as erectile dysfunction and prostate hyperplasia, which have adverse effects on health. Studies have shown that the life expectancy of obese people is 7 years shorter than that of normal people.

Studies have found that Lactobacillus gasseri -BNR17 has the effect of inhibiting the secretion of adiponectin and leptin, which can reduce adipose tissue. Other probiotics, such as Bifidobacterium longum ( B. longum ), Lactobacillus acidophilus ( L. acidophilus ), Pediococcus acidilactici ( P. acidilactici ) and Lactobacillus casei ( L. casei ), have the effect of lowering cholesterol. Lactobacillus casei can reduce the concentration of proinflammatory factors through antioxidant, immunomodulatory and anti-diabetic effects, thereby reducing oxidative stress and inhibiting the production of CD4+T cells. Previous studies on the treatment of obesity have discussed the possibility of translational medicine. According to Larsen et al., obesity is not a disease, but a defect. Although no clear treatment was provided in previous studies, the study found that this "defect" may be adjusted through the connection between intestinal microorganisms and metabolic conditions.

Summary of the invention

The purpose of the present invention is to provide a strain of animal Bifidobacterium lactis subspecies ProSci-246 and products and applications prepared therefrom, so as to provide a new probiotic with good gastrointestinal fluid tolerance, which helps to prevent and/or improve obesity and related diseases caused by obesity.

To achieve the above-mentioned object, the present invention provides a strain of Bifidobacterium animalis subsp. lactis , which is named Bifidobacterium animalis subsp. lactis ProSci-246, and was deposited in the General Microbiology Center of China National Microbiological Culture Collection Committee on March 19, 2024, with a deposit number of CGMCC No. 30074.

Preferably, the Bifidobacterium animalis subsp. lactis is a probiotic, and the gene sequence of its 16S rRNA is shown as SEQ ID NO.1.

SEQ ID NO.1:

ACGGCTCCCCCCACAAGGGTCGGGCCACCGGCTTCGGGTGCTACCCACTTTCATGACTTGACGGGCGGTGTGTACAAGGCCCGGGAACGCATTCACCGCGGCGTTGCTGATCCGCGATTACTAGCGACTCCGCCTTCACGCAGTCGAGTTGCAGACTGCGATCCGAACTGAGACCGGTTTTCAGCGATCCGCCCCACGTCACCGTGTCGCACCGCGTTGTACCGGCCATTGTAGCATGCGTGAAGCCCTGGACGTAAGGG GCATGATGATCTGACGTCATCCCCACCTTCCTCGAGTTGACCCCGGCGGTCCCACATGAGTTCCCGGCATCACCCGCTGGCAACATGCGGCGAGGGTTGCGCT CGTTGCGGGACTTAACCCAACATCTCACGACACGAGCTGACGACGACCATGCACCACCTGTGAACCGGCCCCGAAGGGAAACCGTGTCTCCACGGCGATCCGGCACATGTCAAGCCCAGGTAAGGTTCTTCGCGTTGCATCGAATTAATCCGCATGCTCCGCCGCTTGTGCGGGCCCCCGTCAATTTCTTTGAGTTTTAGCCTTGCGGCCGTACTCCCCAGGCGGGATGCTTAACGCGTTGGCTCCGACACGGGACCCGT GGAAAGGGCCCCACATCCAGCATCCACCGTTTACGGCGTGGACTACCAGGGTATCTAATCCTGTTCGCTCCCCACGCTTTCGCTCCTCAGCGTCAGTGACGGCCC AGAGACCTGCCTTCGCCATTGGTGTTCTTCCCGATATCTACACATTCCACCGTTACACCGGGAATTCCAGTCTCCCCTACCGCACTCCAGCCCGCCCGTACCCGGCGCAGATCCACCGTTAGGCGATGGACTTTCACACCGGACGCGACGAACCGCCTACGAGCCCTTTACGCCCAATAAATCCGGATAACGCTCGCACCCTACGTATTACCGCGGCTGCTGGCACGTAGTTAGCCGGTGCTTATTCGAACAATCCACTCAA CACGGCCGAAACCGTGCCTTGCCCTTGAACAAAAGCGGTTTACAACCCGAAGGCCTCCATCCCGCACGCGGCGTCGCTGCATCAGGCTTGCGCCCATTGTGCA ATATTCCCCACTGCTGCCTCCCGTAGGAGTCTGGGCCGTATCTCAGTCCCAATGTGGCCGGTCACCCTCTCAGGCCGGCTACCCGTCAACGCCTTGGTGGGCCATCACCCCGCCAACAAGCTGATAGGACGCGACCCCATCCCATGCCGCAAAAGCATTTCCCACCCCACCATGCGATGGAGCGGAGCATCCGGTATTACCACCCGTTTCCAGGAGCTATTCCGGTGCACAGGGCAGGTTGGTCACGCATTACTCACCCGTTCG CCACTCTCACCCCGACAGCAAGCTGCCAGGGATCCCGTTCGACTTGCATGTGTTAAGCACGCCGCCAGCGTTCATCCTGAGCCAGTAAACCAACCAAATGC

An application of the live bacteria of animal Bifidobacterium lactis subsp. lactis as described above, wherein the application is an application in the preparation of a drug for preventing and/or improving obesity.

An application of the inactivated bacteria and/or metabolites of Bifidobacterium lactis subsp. animalis as described above, wherein the application is application in the preparation of a drug for preventing and/or improving obesity.

Preferably, the application refers to affecting adult body weight, body fat mass, BMI, waist-to-hip ratio, visceral fat index and health score.

A food and/or health food, drink or medicine containing the above-mentioned animal Bifidobacterium lactis subspecies, wherein the health food, drink or medicine has the function of preventing and/or improving obesity symptoms.

Preferably, the health food, beverage or medicine contains live bacteria, inactivated bacteria and/or metabolites of Bifidobacterium animalis subsp. lactis ProSci-246, wherein the live bacterial count of Bifidobacterium animalis subsp. lactis ProSci-246 is not less than 1×10 ⁶ CFU/mL or 1×10 ⁶ CFU/g.

Therefore, the present invention provides a strain of animal Bifidobacterium lactis subspecies ProSci-246 and the products and applications prepared therefrom, and the specific technical effects thereof are as follows:

(1) The animal Bifidobacterium lactis subspecies ProSci-246 provided by the present invention is a Gram-positive bacterium with an optimal growth temperature of 37°C and an optimal growth pH of 6.0. It has good tolerance to gastrointestinal fluid. After being treated with simulated gastric fluid for 3 hours, the survival rate reached 70.88±0.56%, and after being treated with simulated intestinal fluid for 8 hours, the survival rate reached 88.35±0.54%;

(2) The animal Bifidobacterium lactis subspecies ProSci-246 provided by the present invention can utilize 11 carbohydrates including L-arabinose, D-ribose, D-xylose, D-glucose, methyl-α-D-pyranoglucoside, amygdalin, D-cellobiose, D-lactose, D-melbiose, D-gentiobiose, and D-toulose;

(3) The animal Bifidobacterium lactis subspecies ProSci-246 provided by the present invention has no potential virulence-related genes detected, but contains functional genes related to the production of riboflavin, lactic acid, and exopolysaccharides, the synthesis of bioactive peptides, and acid and bile resistance;

(4) The animal Bifidobacterium lactis subspecies ProSci-246 provided by the present invention can significantly reduce the weight gain and average daily food intake of obese mice, reduce the weight of epididymal fat and inguinal fat in obese mice, alleviate tissue inflammation in obese mice, and reduce fat cell deposition in the liver and fat;

(5) Continuous supplementation of the animal Bifidobacterium lactis subspecies ProSci-246 provided by the present invention for 8 weeks had a significant positive effect on the body weight, body fat mass, BMI, waist-to-hip ratio, visceral fat index and health score of obese adults, and has great application prospects in drugs for preventing and/or improving obesity.

The technical solution of the present invention is further described in detail below through the accompanying drawings and embodiments.

BRIEF DESCRIPTION OF THE DRAWINGS

In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the accompanying drawings required for use in the description of the embodiments of the present invention will be briefly introduced below. Obviously, the accompanying drawings in the following description are only some embodiments of the present invention. For ordinary technicians in this field, other accompanying drawings can be obtained based on these accompanying drawings without paying creative labor.

FIG1 is a circle map of the ProSci-246 genome of Bifidobacterium animalis subsp. lactis obtained in Example 1 of the present invention;

FIG2 is a Gram staining image of Bifidobacterium animalis subsp. lactis ProSci-246 in Example 1 of the present invention;

FIG3 is a graph showing the optimum growth temperature and pH results for Example 1 of the present invention;

FIG4 is a schematic diagram of the settings of the experimental groups in Example 2 of the present invention;

Figure 5 shows the effects of ProSci-246 on weight gain and average daily food intake of obese mice in Example 2 of the present invention, wherein A is the change in weight of mice at 0-20 weeks, B is the weight gain of mice at 0-10 weeks, C is the weight gain of mice at 10-20 weeks, D is the weight gain of mice in the OP group and PO group at 0-10 weeks and 10-20 weeks, E is the average weekly food intake of mice at 0-20 weeks, F is the average daily food intake of mice at 0-10 weeks, G is the average daily food intake of mice at 10-20 weeks, and H is the average daily food intake of mice in the OP group and PO group at 0-10 weeks and 10-20 weeks;

Figure 6 is a graph showing the effects of ProSci-246 on the colon, spleen, and liver of obese mice in Example 2 of the present invention, wherein A is the statistical result of the length of the mouse colon, B is the statistical result of the weight of the mouse spleen, and C is the statistical result of the weight of the mouse liver;

FIG7 is a partial photograph of the effect of ProSci-246 on the colon, spleen and liver of obese mice in Example 2 of the present invention;

FIG8 is a graph showing the effects of ProSci-246 on epididymal fat, inguinal fat, and scapular fat in obese mice in Example 2 of the present invention, wherein A is the statistical result of epididymal fat weight in mice, B is the statistical result of inguinal fat weight in mice, and C is the statistical result of scapular fat weight in mice;

FIG9 is a diagram showing the histological changes of ProSci-246 on colon, spleen, liver, epididymal fat, inguinal fat, and scapular fat in obese mice in Example 2 of the present invention;

Figure 10 is the body composition measurement results of obese adults after 8 weeks of ProSci-246 supplementation in Example 3 of the present invention, where A is the weight change rate, B is the fat mass change rate, C is the body fat percentage change rate, D is the BMI change rate, E is the waist-to-hip ratio change rate, F is the visceral fat index change rate, G is the muscle mass change rate, H is the basal metabolic rate change rate, and I is the health score change rate.

DETAILED DESCRIPTION

The technical solution of the present invention is further described below through the accompanying drawings and embodiments.

In order to make the purpose, technical scheme and advantages of the present application clearer, more thorough and complete, the technical scheme of the present invention is clearly and completely described below through the accompanying drawings and examples. The following detailed descriptions are all descriptions of the embodiments, and are intended to provide further detailed descriptions of the present invention. Unless otherwise specified, all technical terms used in the present invention have the same meaning as those generally understood by those skilled in the art to which the present application belongs.

If no specific experimental steps or conditions are specified in the following examples, the conventional experimental steps or conditions described in the literature in the field can be used. If no manufacturer is specified for the reagents or instruments used, they are all conventional reagent products that can be purchased commercially.

The method for detecting the number of viable bacteria involved in the following embodiments is: adopting the national standard "GB 4789.35-2016 National Food Safety Standard Food Microbiology Detection Lactic Acid Bacteria Detection".

The animal Bifidobacterium lactis subspecies ProSci-246 of the present invention is isolated by the following method:

The collected fecal samples of infants were smeared on site to isolate strains. The specific steps were as follows: first, the samples were diluted in multiples, and 200 μL of the 10 ^-5 and 10 ^-6 dilution gradient samples were evenly smeared on the RCM solid culture medium plate. After evenly smearing, they were placed in an anaerobic tank together with the anaerobic gas production bag and brought back to the laboratory for anaerobically cultured at 37°C in an A35 anaerobic workstation (80% N ₂ , 10% H ₂ , 10% CO ₂ ) for 72 to 96 h. Single colonies with different morphology, size, and color were picked and inoculated into RCM liquid culture medium and anaerobically cultured in a 37°C constant temperature incubator for 24 to 36 h. After the strains grew well, Gram staining and microscopic examination were performed. The isolated strains were preserved, and the genomic DNA of the strains was extracted for subsequent determination and analysis.

The animal Bifidobacterium lactis subspecies ProSci-246 of the present invention forms circular protrusions with a diameter of 1 to 2 mm on the RCM solid culture medium, with irregular edges, rough texture, and milky white opaque colonies.

The animal Bifidobacterium lactis subspecies ProSci-246 of the present invention has the following biological characteristics: the bacteria are short rod-shaped under a microscope, with relatively uniform thickness, and are arranged in single, chain-shaped, uniformly dispersed or clustered states. They have no flagella or cilia and will not move.

The animal bifidobacterium lactis subspecies ( Bifidobacterium animalis subsp. lactis ) ProSci-246 of the present invention has a survival rate of 70.88% after being digested in artificial gastric juice of pH 2.5 for 3 hours, and then continues to be digested in artificial intestinal juice of pH 8.0 for 8 hours, and its survival rate is as high as 88.35%. It has good tolerance to gastrointestinal digestive juice, can enter the human intestine in a living state, and exert health effects. The above characteristics are the basis for being used as probiotics.

Embodiment 1

Isolation and identification of Bifidobacterium animalis subsp. lactis ProSci-246

The Bifidobacterium animalis subsp. lactis ProSci-246 of the present invention is isolated by the following method:

Isolation of Bifidobacterium animalis subsp. lactis ProSci-246

The collected fecal samples of infants were smeared on site to isolate strains. First, the samples were diluted in multiples, and 200 μL of the 10 ^-5 and 10 ^-6 dilution gradient samples were evenly smeared on the RCM solid culture medium plate. After evenly smearing, they were placed in an anaerobic tank together with the anaerobic gas production bag and brought back to the laboratory for anaerobically cultured at 37°C in an A35 anaerobic workstation (80% N ₂ , 10% H ₂ , 10% CO ₂ ) for 72 to 96 h. Single colonies with different morphology, size, and color were picked and inoculated into liquid culture medium and cultured in a 37°C constant temperature incubator for 24 to 36 h. After the strains grew well, Gram staining and microscopic examination were performed. The isolated strains were preserved, and the genomic DNA of the strains was extracted for subsequent determination and analysis.

The components of RCM culture medium are: 10 g peptone, 10 g beef extract powder, 3 g yeast powder, 5 g glucose, 1 g soluble starch, 5 g sodium chloride, 3 g sodium acetate, 0.5 g L-cysteine hydrochloride, 1 L distilled water, adjust the pH to about 6.8, 15 g agar, and sterilize at 121°C for 15 min.

Physiological and biochemical identification, 16S rRNA molecular identification and whole genome sequencing for strain identification

The frozen test strain was inoculated into ML liquid culture medium and cultured at 37°C for 24-36 h. After 2-3 generations of subculture, 30 mL of the bacterial culture at the end of logarithmic growth was placed in a sterile tube for centrifugation. After centrifugation at 8000 g for 5 min (4°C), the bacteria were collected and the supernatant was discarded. The genomic DNA of the strain was extracted using the CTAB freeze-thaw method specifically for lactic acid bacteria.

The components of ML liquid culture medium are: 10 g peptone, 4 g yeast extract powder, 8 g beef extract powder, 20 g glucose, 5 g anhydrous sodium acetate, 2 g triammine citrate, 2 g dimethyl phosphate, 1 mL Tween-80, 0.2 g magnesium sulfate heptahydrate, 0.05 g manganese sulfate tetrahydrate, 1 L distilled water, 0.5 g L-cysteine salt, pH is adjusted to about 6.2, and sterilized at 121°C for 15 min.

The isolated strains were subjected to physiological and biochemical identification and 16S rRNA molecular identification according to the Bergey Manual of Bacterial Identification, using bacterial API50CH (BioMérieux). DNA samples of the strains were extracted, and the samples were subjected to 16S rRNA molecular identification by Shanghai Meiji Biopharmaceutical Technology Co., Ltd. The molecular sequence was compared with NCBI database blastn, confirming that ProSci-246 was animal Bifidobacterium lactis subsp., and its 16S rRNA sequence is shown in SEQ ID NO.1.

The Illumina HiSeq sequencing platform was used to sequence the whole genome of Bifidobacterium animalis subsp. lactis ProSci-246, and the genome circle map is shown in Figure 1. After assembly and alignment, its genome length was 1.95 Mb, and the chromosomal genome contained 1633 coding genes.

c. Colony morphology characteristics

The Bifidobacterium animalis subsp. lactis ProSci-246 of the present invention forms circular protrusions with a diameter of 1 to 2 mm on the RCM solid culture medium, with irregular edges, rough texture, and milky white opaque colonies.

In the clean bench, the purified Bifidobacterium sludge was picked up with an inoculation loop for smearing, and Gram staining was performed after it was dried. The morphology and staining characteristics of the strain were observed and recorded under a microscope. The results are shown in Figure 2. This strain is a Gram-positive bacterium, with a rod-shaped body and various shapes, mostly V-shaped, Y-shaped or baseball-shaped; the colony diameter on the modified MRS solid culture medium is between 1 and 2 mm, the colony edges are neat, creamy white and opaque.

The Bifidobacterium animalis subsp. lactis ProSci-246 of the present invention has the following morphological characteristics: the bacteria are short rod-shaped under a microscope, with relatively uniform thickness, and are arranged in single, chain-shaped, evenly dispersed or clustered states. They do not have flagella or cilia and will not move.

d. Optimal growth temperature and pH

ProSci-246 was inoculated at 2% (v/v) in ML liquid culture medium for activation. The temperature gradient was set at 32, 34.5, 37, 39.5, and 42 °C, and the pH was 6.0. After constant temperature culture for 24 h, the OD value was measured at the absorbance of 600 nm. Three groups of parallels were set for each temperature condition to determine the optimal growth temperature range.

The pH gradient of ML medium was set to 4, 5, 6, 7, 8, and 9. The strains were inoculated into ML liquid medium at 2% (v/v) for activation. The inoculated ML liquid medium was cultured at 37°C for 24 h, and its OD value was measured at 600 nm absorbance. Three groups of parallels were set for each pH condition to determine the optimal pH for strain growth. The results are shown in Figure 3. The optimal growth temperature of ProSci-246 is 37°C, and the optimal growth pH is 6.0.

e. Carbohydrate utilization

ProSci-246 was inoculated into ML liquid culture medium at 2% (v/v), activated and passaged twice at 37°C and pH 6.0, and then centrifuged for 10 min to collect the bacterial sludge, repeatedly rinsed with 0.85% sterile saline and centrifuged until there was no residue. First, fill the small groove in the culture box with sterile distilled water to create a humid environment. Take the above-mentioned bacterial sludge without residue and add it to sterile saline, shake it evenly, and then compare it with the turbidity tube. After the turbidity is consistent, the bacterial liquid is connected to the ampoule tube containing the culture medium, and then the inoculated culture medium is added to the tube of the reagent strip according to the API 50CHL operating instructions, and the tube mouth is sealed with sterile liquid paraffin. Anaerobically cultured at the optimal temperature for 48 h, observe the color change of the reagent strip after culture, and judge the carbohydrate metabolism ability of the strain.

The comparison results show that ProSci-246 can utilize 11 kinds of carbohydrates, namely L-arabinose, D-ribose, D-xylose, D-glucose, methyl-α-D-pyranoglucoside, amygdalin, D-cellobiose, D-lactose, D-myristobiose, D-gentiobiose, and D-toluose.

f. Gastrointestinal fluid tolerance of Bifidobacterium animalis subsp. lactis ProSci-246

Preparation method of simulated gastrointestinal fluid: After sterilizing PBS, adjust the pH value to 2.5 with 1 mol/L HCL, add 3.0 mg/mL pepsin, filter and sterilize with a 0.22 μm microporous filter membrane to prepare simulated artificial gastric juice; after sterilizing PBS, adjust the pH value to 8.0 with 0.1 mol/L NaOH, add 0.1% trypsin and 1.8% ox bile salt, filter and sterilize with a 0.22 μm microporous filter membrane to prepare artificial simulated pancreatic juice.

Gastrointestinal fluid tolerance: The isolated and purified strain was activated and cultured for two generations, and the bacteria were washed twice by centrifugation. The bacteria were collected and 0.5 mL of the re-screened bacterial suspension was added to 4.5 mL of pH 2.5 simulated artificial gastric fluid. The suspension was digested at 37 °C for 3 h. The number of live bacteria was determined by the ML solid culture medium pouring method at 0 h and 3 h, respectively. The survival rate of the strain was calculated using the following formula. Four parallel assays were performed for each sample.

Survival rate = [N1/N0] × 100%

Among them, N0 represents the number of viable bacteria at 0 h; N1 represents the number of viable bacteria after 3 h of digestion in simulated gastric juice.

Afterwards, 0.5 mL of artificial gastric juice containing bacteria that had been digested for 3 h was added to 4.5 mL of artificial intestinal juice and cultured in a 37°C water bath. The number of live bacteria was determined by the ML solid culture medium pouring method after 0 h and 8 h. The survival rate of the strain was calculated using the following formula. Four parallels were made for each sample.

Survival rate = [N1/N0] × 100%

Among them, N0 represents the number of viable bacteria at 0 h; N1 represents the number of viable bacteria after 8 h of digestion with simulated pancreatic juice.

Tolerance effect: After animal Bifidobacterium lactis subspecies ProSci-246 was treated with simulated gastric fluid and simulated intestinal fluid, the results are shown in Table 1 below.

Table 1 Survival of Bifidobacterium animalis subsp. lactis ProSci-246 in simulated gastrointestinal digestive fluid

;

As shown in Table 1, animal Bifidobacterium lactis subspecies ProSci-246 has good tolerance characteristics, and the survival rate can reach 88.35% after 8 h of treatment with simulated intestinal fluid.

According to the gastrointestinal fluid tolerance effect of this example, it can be seen that animal Bifidobacterium lactis subsp. ProSci-246 has a good tolerance to gastrointestinal fluid in the intestine and has probiotic properties.

g. Detection of probiotic-related genes of Bifidobacterium animalis subsp. lactis ProSci-246

The animal Bifidobacterium lactis subsp. lactis ProSci-246 was compared with the VFDB database (Virulence Factors of Pathogenic Bacteria) and the VirulenceFinder database (https://cge.cbs.dtu.dk/services/VirulenceFinder), with sequence similarity > 90% and sequence coverage > 60% as screening criteria. The results showed that no potential virulence-related genes were detected in the ProSci-246 genome.

Genes related to probiotic functions were detected in the ProSci-246 genome of Bifidobacterium animalis subsp. lactis and annotated to obtain functional genes related to the production of riboflavin, lactic acid, and extracellular polysaccharides, the synthesis of bioactive peptides, and acid and bile resistance. The specific genes are shown in Table 2.

Table 2 Probiotic-related genes

;

Embodiment 2

The effect of animal Bifidobacterium lactis subspecies ProSci-246 on obesity in mice was investigated as follows:

The mice were randomly divided into 5 groups, and the treatment methods of each group were as follows:

NC group (SPF 4-week-old male C57BL/6J mice, weighing 20±2 g), 16 mice, fed with saline for 0-20 weeks); OB group (SPF 4-week-old C57BL/6JGpt-Chr1 ^YP1 mice, weighing 20±2 g), 16 mice, fed with saline for 0-20 weeks); OP group (SPF 4-week-old C57BL/6JGpt-Chr1 ^YP1 mice, weighing 20±2 g), 16 mice, fed with saline for 0-10 weeks and ProSci-246 probiotic powder for 10-20 weeks); PO group (SPF 4-week-old C57BL/6JGpt-Chr1 ^YP1 mice, weighing 20±2 g) 16 mice, fed with ProSci-246 probiotic powder from 0 to 10 weeks, and fed with normal saline from 10 to 20 weeks); PP group (SPF 4-week-old C57BL/6JGpt-Chr1 ^YP1 mice, weight 20±2 g) 16 mice, fed with ProSci-246 probiotic powder from 0 to 20 weeks), the probiotic feeding amount was 4×10 ⁹ CFU/d. The schematic diagram of the treatment settings of each group is shown in Figure 4.

C57BL/6JGpt-Chr1 ^YP1 mice are a chromosome replacement mouse strain that exhibits a spontaneously obese phenotype on a regular diet and can be purchased commercially.

Preparation method of animal Bifidobacterium lactis subsp. ProSci-246 probiotic powder: freeze-dry the bacterial sludge without residue obtained in e of Example 1, grind it, and mix it evenly with oligofructose and polydextrose in a mass ratio of 1:9:9 to obtain the powder.

Experimental Results

(1) Effects of ProSci-246 on weight gain and average daily food intake in obese mice

The results are shown in Figure 5. There was no significant difference in the initial body weight of mice in each group. At 10 weeks, the body weight of mice in the OB and OP groups exceeded that of the NC group by 20%, and the body weight gain of mice in the PP and PO groups was significantly lower than that of the OB group ( P < 0.01) and the OP group ( P < 0.05). The body weight of mice in the OB group was significantly higher than that of the OP group from the 14th week ( P < 0.05). At 20 weeks, the body weight gain of mice in the OB group was significantly higher than that of the OP group ( P < 0.0001) and the PP group ( P < 0.001), and the body weight gain of mice in the PO group was significantly higher than that of the OP group ( P < 0.001) and the PP group ( P < 0.01).

From 0 to 10 weeks, the average daily food intake of mice in the OB group was not significantly different from that in the NC, OP, PO and PP groups; the average daily food intake of mice in the OP and PP groups was significantly different ( P < 0.05). From 10 to 20 weeks, the average daily food intake of mice in the OB group was significantly different from that in the NC and PP groups ( P < 0.05).

(2) Effect of ProSci-246 on epididymal fat and inguinal fat weight in obese mice

As shown in Figures 6 and 7, supplementation with Bifidobacterium lactis ProSci-246 had no significant effect on the colon length of mice; the liver weight of mice in the OB group was significantly higher than that in the NC group ( P < 0.001), and there was no significant difference in the liver weight of mice among the other four groups; there was no significant effect on the spleen weight of mice in each group.

As shown in Figure 8, the weight of epididymal fat and inguinal fat in the OB group was significantly heavier than that in the NC group ( P <0.0001), PO group ( P <0.01) and PP group ( P <0.001), and there was no significant difference among the OP group, PO group and PP group. The weight of scapular fat in the OB group was significantly heavier than that in the NC group ( P <0.01), and there was no significant difference among the OB group, OP group, PO group and PP group.

(3) Effect of ProSci-246 on tissue inflammation and adipocyte deposition in liver and adipose tissues of obese mice

As shown in Figure 9, the results showed that the colon: the crypt structure of the intestinal tissue was destroyed in the OB group of mice, and there was mild to moderate inflammatory cell infiltration in the colon of the OP, PO and PP group of mice.

Spleen: A small amount of adipocyte formation was observed in the spleens of mice in the OB, OP, PO and PP groups. A small amount of inflammatory cell infiltration was observed in the OB group, a very small amount of inflammatory cell infiltration was observed in the OP and PO groups, and no obvious pathological changes were observed in the PP group.

Liver: In the OB group, large areas of ballooning degeneration, fatty degeneration of a large number of cells with moderate inflammatory cell infiltration were observed; in the OP, PO and PP groups, mild ballooning degeneration, fatty degeneration of a small number of cells with a small amount of inflammatory cell infiltration were observed.

Epididymal fat: some adipocytes were ruptured and stacked, and a small number of adipocytes were atrophied in the OB, OP, and PO groups; a small amount of inflammatory cell infiltration was observed in the OB and PO groups; and a very small amount of inflammatory cell infiltration was observed in the OP and PP groups.

Inguinal fat: some adipocytes were ruptured and stacked, and a small number of adipocytes were atrophied in the OB, OP, and PO groups; very small amounts of inflammatory cell infiltration were observed in the OB, OP, PO, and PP groups.

Scapular fat: Very little inflammatory cell infiltration was observed in the OB, OP, PO, and PP groups.

Embodiment 3

The effect of animal Bifidobacterium lactis subspecies ProSci-246 on human obesity was investigated as follows:

The basic requirements for volunteers to participate in the trial are as follows:

① Male or female, aged between 18 and 65 years old; ② Body mass index (BMI) ≥ 28 kg/m ² ; ③ Male waist circumference ≥ 90 cm, female waist circumference ≥ 85 cm; ④ No use of antibiotics, probiotics or anti-obesity drugs in the month before the trial; ⑤ No history of chronic gastrointestinal diseases; ⑥ Agree to participate in this study and sign a written informed consent.

The trial plan was based on the "Chinese Expert Consensus on Medical Nutritional Therapy for Overweight/Obesity (2016 Edition)" and individualized diet plans and exercise recommendations were formulated according to the subject's body composition.

Complete data of 90 volunteers were obtained, including 43 in the probiotic group and 47 in the placebo group. Volunteers in the probiotic group took 2g of animal Bifidobacterium lactis subsp. ProSci-246 probiotic powder every day (the number of live bacteria of ProSci-246 was 3×10 ¹⁰ CFU). The placebo group only took the same amount of oligofructose and polydextrose (excluding live ProSci-246) as the volunteer group every day. The body composition of obese adults (including weight, body fat mass, body fat percentage, BMI, waist-to-hip ratio, visceral fat index, muscle mass, and health score) was measured at the 4th and 8th weeks of the trial.

The preparation method of animal Bifidobacterium lactis subspecies ProSci-246 probiotic powder is the same as that in Example 2.

The results are shown in Figure 10. Compared with the placebo group, continuous supplementation of ProSci-246 for 4 weeks significantly reduced the weight, body fat mass and BMI of obese adults. After 8 weeks of supplementation, the waist-to-hip ratio and visceral fat index were significantly reduced. There was no significant effect on the rate of change of muscle mass, and the health score was significantly improved.

Therefore, the animal Bifidobacterium lactis subspecies ProSci-246 provided by the present invention is a Gram-positive bacterium, has an optimal growth temperature of 37°C, an optimal growth pH of 6.0, has good tolerance to gastrointestinal fluid, can utilize 11 types of carbohydrates, has no detected potential virulence-related genes, and contains functional genes related to good health; can significantly reduce the weight gain and average daily food intake of obese mice, reduce the weight of epididymal fat and inguinal fat in obese mice, reduce tissue inflammation in obese mice, and reduce fat cell deposition in the liver and fat; has a significant positive effect on the weight, body fat mass, BMI, waist-to-hip ratio, visceral fat index and health score of obese adults, and has great application prospects in products for preventing and/or improving obesity.

Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that they can still modify or replace the technical solution of the present invention with equivalents, and these modifications or equivalent replacements cannot cause the modified technical solution to deviate from the spirit and scope of the technical solution of the present invention.

Claims (6)

1. The bifidobacterium animalis subspecies (Bifidobacterium animalis subsp. Lactis) are named as ProSci-246 and are preserved in the China general microbiological culture Collection center (CGMCC) with the preservation number of 30074, and the bifidobacterium animalis subspecies are probiotics, and the gene sequence of 16S rRNA is shown as SEQ ID NO. 1.

2. Use of a viable bifidobacterium animalis subsp lactis as claimed in claim 1 wherein: the application is the application in preparing the medicine for improving obesity.

3. A food product comprising the bifidobacterium animalis subspecies lactis of claim 1.

4. A health food comprising the bifidobacterium animalis subspecies lactis of claim 1, characterized in that: the health food has the function of improving obesity; the health food contains live bacteria of animal bifidobacterium subspecies ProSci-246, wherein the number of live bacteria of animal bifidobacterium subspecies ProSci-246 is not lower than 1X 10 ⁶ CFU/mL or 1X 10 ⁶ CFU/g.

5. A beverage comprising the bifidobacterium animalis subspecies lactis of claim 1, characterized in that: the beverage has obesity improving effect; the beverage contains live bacteria of animal bifidobacterium subspecies ProSci-246, wherein the number of live bacteria of animal bifidobacterium subspecies ProSci-246 is not lower than 1X 10 ⁶ CFU/mL or 1X 10 ⁶ CFU/g.

6. A pharmaceutical product comprising the bifidobacterium animalis subspecies lactis of claim 1, characterized in that: the medicine has the function of improving obesity; the medicine contains live bacteria of animal bifidobacterium subspecies ProSci-246, wherein the number of the live bacteria of animal bifidobacterium subspecies ProSci-246 is not lower than 1X 10 ⁶ CFU/mL or 1X 10 ⁶ CFU/g.