CN113403225B - Bifidobacterium animalis NSY0201 and application thereof in preparation of health care products or medicines for enhancing immunity of organisms - Google Patents
Bifidobacterium animalis NSY0201 and application thereof in preparation of health care products or medicines for enhancing immunity of organisms Download PDFInfo
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- CN113403225B CN113403225B CN202110622102.7A CN202110622102A CN113403225B CN 113403225 B CN113403225 B CN 113403225B CN 202110622102 A CN202110622102 A CN 202110622102A CN 113403225 B CN113403225 B CN 113403225B
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Abstract
The invention discloses bifidobacterium animalis NSY0201 and application thereof in preparing health products or medicines for enhancing immunity of organisms. The Bifidobacterium animalis NSY0201 is named as Bifidobacterium animalisBifidobacterium animalisThe protein is preserved in China center for type culture Collection, has the preservation number of CTCC M2021629, has the nucleotide sequence shown in SEQ ID No.1, and has the function of high-yield extracellular polysaccharide. The bifidobacterium animalis NSY0201 and the freeze-dried powder thereof can achieve the effects of enhancing the immune function and disease resistance of organisms by inhibiting DNFB-induced DTH, improving the serum hemolysin level, phagocytic index and NK cell activity in mice and accelerating the proliferation capacity of lymphocytes, are safe to use, can be used for preparing health-care products or medicines, and have good application prospects.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to bifidobacterium animalis NSY0201 and application thereof in preparation of health-care products or medicines for enhancing immunity of organisms.
Background
Immunity is the body's own defense mechanism, and is the body's ability to recognize and destroy any foreign body (virus, bacteria, etc.) that invades from the outside, to treat aged, damaged, dead, denatured self cells, and to recognize and treat mutant cells and virus-infected cells in the body. Human beings can live in dangerous environments for millions of years, and mainly rely on self resistance.
The body with low immunity is easy to be infected or suffer from cancer, the weak resistance of children can cause poor physical and intelligence development, serious diseases are easy to be induced, and the health problems which are caused are repeated. Therefore, modern people increasingly attach importance to the improvement of autoimmunity. At present, diet conditioning, exercise enhancement, zinc supplement and the like are mostly adopted for enhancing the immunity, but the effects of the above are slow.
Probiotics are active microorganisms which are beneficial to a host and can change the composition of flora at a certain part of the host by colonizing in a human body, and can generate the effect of benefiting the health of the organism. At present, probiotics have important functions in the aspects of enhancing the immune function, resisting tumors and the like. Therefore, the search for new probiotics for continuously enhancing the immunity of the organism is very beneficial to the physical and mental health of patients.
Disclosure of Invention
The invention aims to provide bifidobacterium animalis NSY0201 and application thereof in preparing health-care products or medicines for enhancing immunity of organisms. The Bifidobacterium animalis NSY0201 has safe source, and has functions of enhancing organism immunity and resisting disease.
In order to realize the purpose of the invention, the invention adopts the following technical scheme to realize:
the invention provides bifidobacterium animalis NSY0201, and the classification name of the bifidobacterium animalis NSY0201 is bifidobacterium animalisBifidobacterium animalisAnd the product is preserved in China center for type culture Collection with the preservation number of CTCC M2021629.
Further, the nucleotide sequence of the bifidobacterium animalis NSY0201 is shown as SEQ ID No. 1.
Further, the colony of the bifidobacterium animalis NSY0201 is milky white to light yellow, round or quasi-round and has the diameter of 1-2 mm; the surface is raised, smooth and dull, and the edge is irregularly wavy; the thallus is in a short rod shape, the two ends are blunt and round, the thallus is fresh and forked, the surface is smooth, and no spores exist.
Further, the concentration of the extracellular polysaccharide produced by the bifidobacterium animalis NSY0201 is 306 mg/L.
The invention also provides application of the bifidobacterium animalis NSY0201 in preparing health care products or medicines for enhancing the immunity of organisms.
Furthermore, the health care product or the medicine contains live bacteria with the content of not less than 4 multiplied by 1011 CFU/g Bifidobacterium animalis NSY0201 freeze-dried powder.
Furthermore, the dosage of the bifidobacterium animalis NSY0201 freeze-dried powder is 0.2 to 5 percent of the mass of the health care product or the medicine.
Further, the preparation method of the bifidobacterium animalis NSY0201 freeze-dried powder comprises the following steps: activating the bifidobacterium animalis NSY0201, inoculating the bifidobacterium animalis NSY0201 in an improved MRS liquid culture medium in an inoculation amount of 2-5% (v/v), culturing at 37 ℃ for 14-18 h, continuously culturing for 2 generations of expansion culture, centrifuging and cleaning the obtained fermentation liquor at low temperature, uniformly mixing the collected bacterial sludge and a freeze-drying protective agent in a ratio of 1:1.5-2, fully emulsifying, and performing vacuum freeze-drying to obtain the bifidobacterium animalis NSY0201 freeze-dried powder.
Further, the bifidobacterium animalis NSY0201 can improve the serum hemolysin level, the number of hemolytic plaques and the NK cell activity in animals.
Furthermore, the bifidobacterium animalis NSY0201 can inhibit DNFB-induced DTH and accelerate the proliferation capacity of lymphocytes, thereby achieving the effect of enhancing the body immunity.
Compared with the prior art, the invention has the following advantages and technical effects:
1. the animal bifidobacterium NSY0201 is separated from the natural fermented cheese, has safe sources and high extracellular polysaccharide yield, can use various carbohydrates as growth energy sources, and has wide growth conditions; has strong acid and choline tolerance, and is suitable for survival in gastrointestinal tract.
2. Bifidobacterium animalis NSY0201 and freeze-dried powder prepared by the same can inhibit or relieve DTH generated by DNFB induction, improve mononuclear phagocyte system function, improve antibody generating capacity and the like by increasing the weight difference of about ears of DNFB-induced DTH, improving serum hemolysin level, hemolytic plaque number, phagocytic index and NK cell activity in a mouse and accelerating the proliferation capacity of lymphocytes, and finally play a role in enhancing the immune function and disease resistance of an organism. Provides a theoretical basis for developing novel health care products or medicines for enhancing the immunity of the organism.
Drawings
FIG. 1 is a photograph of Bifidobacterium animalis NSY0201 in plate culture.
FIG. 2 is a microscopic photograph of Bifidobacterium animalis NSY 0201.
FIG. 3 is a photograph of fermentation broth of Bifidobacterium animalis NSY 0201.
Detailed Description
The technical solution of the present invention will be further described in detail with reference to the following specific examples. In the following examples, unless otherwise specified, the experimental methods used were all conventional methods, and materials, reagents and the like used were all available from biological or chemical reagents companies.
Example 1: isolation and identification of Bifidobacterium animalis NSY0201
1. Isolation and screening of strains
Natural fermented cheese from the inner Mongolia Cinera Lingii was used as a sample. Adding 3 samples into 10% skimmed milk culture medium according to the amount of 1%, culturing at 37 deg.C, curdling, diluting gradually to 10%-8And (4) gradient. Selecting 3 appropriate dilutions, respectively taking 1mL of the diluted solutions in an improved MRS solid culture medium, carrying out anaerobic culture at 37 ℃ for 48h, carrying out colony morphology observation, selecting single colonies with different morphologies, carrying out streak separation on the improved MRS solid culture medium again, carrying out anaerobic culture at 37 ℃ for 48h, repeating the streak separation for 3 generations to obtain purified strains, carrying out gram staining and microscopic observation, selecting strains which are positive in gram staining and are spherical, rod-shaped or polymorphic in microscopic observation, and screening 6 strains which meet the requirements in total, wherein the number of the strains is NS 001-006 NS. The 6 strains are subjected to anaerobic amplification culture by using an improved MRS liquid culture medium, and the obtained bacterial liquid is preserved by using 30% of glycerolIn a refrigerator at-80 ℃.
And (3) centrifuging the bacterial liquid of the screened 6 strains, and finding that the bacterial liquid of the NS003 cannot form a solid precipitate after centrifugation, and bacterial mud is loose, possibly because the NS003 generates more extracellular polysaccharide. The 6 strains were negatively stained by negative staining, and the capsular staining of NS003 was found to be the thickest.
The content of extracellular polysaccharide is measured by a phenol-sulfuric acid method in the experiment, and then the content is measured by a colorimetric method. The results shown in Table 1 show that the yield of exopolysaccharide of NS003 is the highest, 306 mg/L, which is much higher than that of other 5 strains. NS003 was therefore selected for further testing and was relabeled as NSY 0201.
Table 1: exopolysaccharide production of the strain
| Serial number | Exopolysaccharide production (mg/L) |
| NS001 | 117 |
| NS002 | 168 |
| NS003 | 306 |
| NS004 | 135 |
| NS005 | 124 |
| NS006 | 154 |
2. Morphological characteristics
Morphological observation of strain NSY0201 resulted in gram positive strain NSY0201 with milky white to yellowish, round or quasi-round, 1-2mm diameter, as shown in fig. 1 and 2; the surface is raised, smooth and dull, and the edge is irregularly wavy; the thallus is in a short rod shape, the two ends are blunt and round, the thallus is fresh and forked, the surface is smooth, and no spores exist.
3. 16S rRNA identification of strains
DNA of strain NSY0201 was extracted as a template, amplified using bacterial 16S rRNA universal primers 27F (5'-GAG AGT TTG ATC CTG GCT CAG-3') and 1492R (5'-ACG GAT ACC TTG TTA CGA CTT-3'), and the amplified fragments were sent to Bio-Inc for sequencing, the sequencing results are shown in SEQ ID No. 1. BLAST alignment showed that strain NSY0201 andBifidobacterium animalisthe sequence similarity of (a) was 99.79%, and therefore, it was judged that the strain NSY0201 was bifidobacterium animalis.
The strain NSY0201 is selected for strain preservation, and the preservation unit of bifidobacterium animalis NSY0201 is as follows: china Center for Type Culture Collection (CCTCC); address: eight Lopa in Wuchang region of Wuhan city, Hubei province; the preservation date is as follows: 31/5/2021; bifidobacterium animalisBifidobacterium animalisThe accession number of (1) is CTCC M2021629.
Example 2: physiological and biochemical properties of bifidobacterium animalis NSY0201
Activating the preserved Bifidobacterium animalis NSY0201, inoculating to modified MRS culture medium at 3%, culturing at 37 deg.C for 16h, continuously culturing for 3 generations, and performing amplification culture to obtain fermentation broth (FIG. 3) for performing various physiological and biochemical characteristics experiments. The formula of the improved MRS culture medium is as follows: 18 g of glucose, 10g of peptone, 7g of beef extract, 4 g of yeast extract, 2g of diammonium hydrogen citrate, 5g of sodium acetate, 2g of dipotassium hydrogen phosphate, 0.58 g of magnesium sulfate, 0.25 g of manganese sulfate, 801.0 ml of tween-801.0, 0.5g of L-cysteine hydrochloride, 1000ml of distilled water and pH =6.8 +/-0.5.
1. Routine physiological and biochemical experiments
The fermentation liquor is used for carrying out methyl red experiments, acetyl methyl methanol experiments, nitrate reduction experiments and arginine ammonia production experiments. The results are shown in table 2, bifidobacterium animalis NSY0201 does not produce nitrate reductase and can not reduce nitrate to harmful nitrite; arginine cannot be decomposed to produce ammonia gas.
Table 2: results of routine physiological and biochemical experiments
| Bacterial strains | Methyl Red | Acetylmethylmethanol | Nitrate reduction | Production of ammonia from arginine |
| NSY0201 | + | + | - | - |
2. Carbohydrate metabolism
And (4) carrying out metabolism detection on various carbohydrates by using the fermentation liquor. The results are shown in Table 3, Bifidobacterium animalis NSY0201 can utilize 15 of the 16 carbon sources listed, and the only thing that can not be utilized is mannitol, so the growth conditions of the bacterium are wide.
Table 3: carbohydrate metabolism results
| Bacterial strains | Arabinose | Xylose | Ribose | Glucose | Mannose | Fructose | Galactose | Sucrose |
| NSY0201 | + | + | + | + | + | + | + | + |
| Bacterial strains | Maltose | Cellobiose | Lactose | Mannitol | Sorbitol | Cotton seed candy | Starch | Trehalose |
| NSY0201 | + | + | + | - | + | + | + | + |
3. Determination of the growth Curve of the Strain
Activating the preserved bifidobacterium animalis NSY0201, inoculating the activated bifidobacterium animalis NSY0201 into an improved MRS culture medium in an inoculation amount of 3%, anaerobically culturing at 37 ℃, sampling every 2 hours, and measuring the OD (optical density) value of strain fermentation liquor at the position of 600nm of wavelength at different culture times.
As shown in Table 4, Bifidobacterium animalis NSY0201 was cultured until 6-16h entered the late logarithmic growth phase, and 16-20h was the stationary phase of growth.
Table 4: OD value of growth curve
| Time | OD value |
| 0 | 0.225 |
| 2 | 0.528 |
| 4 | 0.621 |
| 6 | 0.758 |
| 8 | 1.00 |
| 10 | 1.26 |
| 12 | 1.60 |
| 14 | 1.95 |
| 16 | 2.47 |
| 18 | 2.52 |
| 20 | 2.52 |
4. Gastric juice resistance test
Adding pepsin 3.0g/L into sterilized PBS with pH values of 2.0, 2.5 and 3.0 by filtration sterilization method to obtain artificial gastric juice; adding 100 muL fermentation liquor into 900 muL simulated gastric juice, and placing for 0.5h, 1.0h and 1.5h under an anaerobic condition at 37 ℃; and adding 900 muL sterile physiological saline into 100 muL bacterial suspension as an experimental control. Sampling, diluting and coating a flat plate, counting viable bacteria, and calculating the survival rate.
The results are shown in table 5, and bifidobacterium animalis NSY0201 can resist strong acidity, so that the bifidobacterium animalis NSY0201 has better tolerance to gastric juice and is suitable for survival and colonization in the stomach.
Table 5: gastric juice resistance test results
| Survival rate (%) | 2.0 | 2.5 | 3.0 |
| 0.5 | 86.54 | 73.65 | 62.44 |
| 1.0 | 80.03 | 68.33 | 59.47 |
| 1.5 | 70.12 | 63.18 | 52.13 |
5. Bile salt tolerance test
3.0g/L of bovine bile salt and 2.0g/L of sodium thioglycolate were added to the MRS liquid culture medium. Bifidobacterium animalis NSY0201 was inoculated into MRS liquid medium containing bile salts and anaerobically cultured at 37 deg.C, and the control group was MRS liquid medium. Sampling, diluting and coating the sample on a flat plate, counting viable bacteria, and calculating the survival rate.
The results are shown in table 6, bifidobacterium animalis NSY0201 is well tolerated by bile salts and is therefore able to colonise and survive in the biliary tract.
Table 6: results of bile salt tolerance experiments
| 1h | 2h | 3h | 4h | 5h | 6h | |
| Survival rate (%) | 88.02 | 79.52 | 68.02 | 61.00 | 53.84 | 45.33 |
Example 3: preparation of bifidobacterium animalis NSY0201 freeze-dried powder
Streaking preserved Bifidobacterium animalis NSY0201 on modified MRS solid plate, culturing at 37 deg.C for activation, selecting single colony, inoculating to modified MRS liquid culture medium, culturing at 37 deg.C for 16 hr, and inoculating to culture medium at 3%Inoculating the strain to an improved MRS culture medium, culturing at 37 ℃ for 16h, and continuously culturing for 2 generations for amplification culture; the obtained fermentation liquor is centrifuged at 12000r/min at 4 ℃ for 10min, supernatant is discarded, bacterial sludge is collected, after the bacterial sludge is washed for 3 times by using ultrapure water, the bacterial sludge and a freeze-drying protective agent (the formula is 30g of trehalose, 10g of skim milk powder, 7g of inulin, 0.5g of isomaltooligosaccharide, 0.2g of glycine, 1g of tween 80 and 1000mL of pure water) are mixed according to the mass ratio of 1:1.7, and after full emulsification, freeze-drying in vacuum is carried out to obtain the freeze-dried live bacterial powder of the animal bifidobacterium NSY 0201. And (4) pouring and counting the freeze-dried viable bacteria powder, and storing the viable bacteria powder in a refrigerator at the temperature of 18 ℃ below zero in vacuum for later use. The strain content of lyophilized live powder of Bifidobacterium animalis NSY0201 is 4 × 1011 CFU/g。
Example 4: experimental study on function of bifidobacterium animalis NSY0201 in enhancing immunity
1. Grouping and dose selection
200 Kunming mice were purchased and acclimatized for one week, and then divided into 4 groups: placebo, 0.2g, 1.0g and 5.0g dose groups, 50 per group, were individually gavaged. Wherein, the mice of the blank control group are subjected to intragastric lavage by 0.2mL of distilled water; dose groups 0.2g, 1.0g, 5.0g of the lyophilized powder of Bifidobacterium animalis NSY0201 prepared in example 3 was diluted with 100g of ultrapure water, and then the diluted solution equivalent to distilled water was used to perfuse the stomach of the mice.
2. Detailed description of the invention
The stomach is drenched once a day for 30 days continuously according to the dose of each group, and then each immune index is measured.
(1) Delayed allergic response (DTH) assay in mice
Delayed allergic test-degree of ear swelling. And (3) performing intragastric administration at 26d, depilating the abdomen of each mouse by about 4 cm multiplied by 4 cm by using a depilatory cream, uniformly coating 50 mu L of DNFB solution on the depilated part for sensitization, uniformly coating 10 mu L of DNFB solution on the two sides of the right ear of the mouse after 5 d, killing the mouse after 24 h, shearing the left ear shell and the right ear shell, taking a circular sheet with the diameter of 6 mm by using a puncher, weighing, and calculating the weight difference of the left ear and the right ear.
The results are shown in table 7, and in the DNFB-induced DTH experiment, the difference between the weights of the left and right ears of the 0.2g dose group, the 1.0g dose group and the 5.0g dose group was significantly higher than that of the blank control group (P < 0.05), indicating that DTH appears in the mice after DNFB treatment, and the experimental results of the dose groups were positive. The delayed allergy belongs to cell immunity, is a hypersensitivity mediated by specific sensitized effector T cells, and positive results also indicate that the animal bifidobacterium NSY0201 can inhibit or relieve DTH (glutathione) generated by DNFB induction and improve the body immunity of mice.
Table 7: results of DTH measurement
| Grouping | Weight difference between right and left ears |
| Blank control group | 1.85±0.25 |
| 0.2g dose group | 2.76±0.58 |
| 1.0g dose group | 2.86±0.45* |
| 5.0g dose group | 2.99±0.61* |
(2) Canavastin A (Con A) Induction of mouse spleen lymphocyte transformation experiment (MTT method)
Aseptically collecting spleen from each group of mice on 26 th day after intragastric administration, preparing spleen lymphocyte suspension, and adjusting spleen cell concentration to 3 × 10 with RPMI 1640 complete culture solution6 CFU/mL, splenic lymphocyte proliferative responses according to the in-program MTT method, in which a portion of cells were induced with Con A: (Con A+) Non-induced cells were used as a control (Con A)-) Finally, the absorbance value (A) is measured at a wavelength of 570nm, and Con A is calculated+And Con A-Absorbance difference for each well. Results from each dose group were compared to a solvent control group for analysis of variance.
The results are shown in Table 8, in the Con A-induced splenic lymphocyte transformation experiments of mice, the difference of the absorbance values of the Con A wells and the Con A wells of each dose group is higher than that of the blank control group (P < 0.05), and the bifidobacterium animalis NSY0201 is proved to be capable of accelerating the proliferation of lymphocytes.
Table 8: con A induced mouse spleen lymphocyte transformation experimental results
| Grouping | Con A+And Con A-Difference in light absorption value of |
| Blank control group | 0.199±0.043 |
| 0.2g dose group | 0.251±0.042 |
| 1.0g dose group | 0.303±0.065* |
| 5.0g dose group | 0.298±0.076* |
(3) Serum hemolysin assay
The mice were injected intraperitoneally with 20% SRBC (0.2 ml/mouse) on the 25 th day of gavage, and the mice were subjected to blood sampling by eye picking on the 5 th day after immunization, and serum was separated and subjected to serum hemolysin assay on a microaerogram: incubating for 3h at 37 ℃, counting the hemagglutination degree, and calculating the corresponding antibody product number.
The results are shown in Table 9, and the half hemolysis value HC of each dose group of mice50Is obviously higher than a blank control group (P is less than 0.01), and proves that the bifidobacterium animalis NSY0201 can improve the content of serum hemolysin in a mouse body, and further can improve the function of a mononuclear phagocyte system, namely the disease resistance of the mouse.
Table 9: results of serum hemolysin assay
| Serum hemolysin (%) HC50 | |
| Blank control group | 70.895±10.368 |
| 0.2g dose group | 78.758±14.145 |
| 1.0g dose group | 80.053±14.286* |
| 5.0g dose group | 81.361±15.027* |
(4) Antibody producing cell (PFC) assay
Mice were injected intraperitoneally with 20% SRBC (0.2 ml/mouse) on day 25 of gavage, sacrificed on day 5 after immunization, spleen was dissected and prepared for splenocyte suspension, and the spleen was used with RPMI 1640The whole culture solution can adjust the concentration of splenocytes to 3 × 106 CFU/mL, agarose slides prepared by procedure, in CO2Incubator (37 ℃, 5% CO)2) After 1.5h incubation, complement was added and further incubated for 1.5h, and the number of lyso-plaques formed on each agar thin-layer slide was counted.
The results are shown in table 10, and the antibody-producing cell detection experiment shows that the number of hemolytic plaques caused by the same number of splenocytes of mice in each dose group is higher than that of mice in a blank control group (P < 0.05), which indicates that the bifidobacterium animalis NSY0201 significantly enhances the function of B lymphocytes to produce corresponding antibodies, namely, enhances the humoral immune response function of the body.
Table 10: result of PFC detection
| Number of hemolytic plaques/103Spleen cell | |
| Blank control group | 20.065±4.778 |
| 0.2g dose group | 23.004±3.989 |
| 1.0g dose group | 25.678±3.802* |
| 5.0g dose group | 25.649±3.696* |
(5) Carbon clearance test of mice
Each group of mice was injected with 0.1ml/10g of India ink (4-fold dilution) into the tail vein in order. Collecting 20 μ l blood of each mouse at 2min and 10min after injecting Chinese ink, quickly adding into 2ml 0.1% sodium carbonate solution, shaking, and measuring absorbance at 600nm with ultraviolet-visible spectrophotometer; and weighing the liver and spleen of the mouse, and calculating the phagocytosis index according to a formula. Results from each dose group were compared to a solvent control group for analysis of variance.
The results are shown in table 11, and carbon clearance experiments show that the phagocytic index of mice in each dose group is higher than that of mice in a blank control group (P is less than 0.05), which indicates that the animal bifidobacterium NSY0201 can improve the capability and speed of phagocytic carbon granules of macrophages, and further improve the immune function of the body.
Table 11: results of phagocytic index in mouse carbon clearance assay
| Phagocytic index in clearance test | |
| Blank control group | 4.2788±0.8945 |
| 0.2g dose group | 4.5820±0.5498 |
| 1.0g dose group | 6.3894±0.5482* |
| 5.0g dose group | 6.4698±0.7794* |
(6) NK cell Activity assay
Immediately after blood is taken from mouse eyeballs, blood is centrifuged at 4000 r/min for 10min to obtain serum, and the activity of NK cells is measured according to the measuring method of a Lactate Dehydrogenase (LDH) kit.
The result is shown in table 12, and the NK cell activity measurement finds that the NK cell activity of mice in each dose group is higher than that of mice in a blank control group (P is less than 0.05), so that the lactobacillus plantarum NSY0201 can obviously improve the NK cell activity of the mice, and then the whole immunity of a body can be improved.
Table 12: NK cell activity assay structure
| NK cell Activity (%) | |
| Blank control group | 32.745±5.395 |
| 0.2g dose group | 39.762±6.398 |
| 1.0g dose group | 42.390±6.221* |
| 5.0g dose group | 43.933±6.684* |
In summary, the animal experiments prove that the bifidobacterium animalis NSY0201 and the freeze-dried powder thereof can inhibit or relieve DTH generated by DNFB induction, improve the mononuclear phagocyte system function, improve the antibody generating capacity and the like by increasing the weight difference of the DNFB-induced DTH, improving the serum hemolysin level, the number of hemolytic plaques, the phagocytic index and the NK cell activity in a mouse and accelerating the proliferation capacity of lymphocytes, and finally play a role in enhancing the immune function and the disease resistance of an organism. Also, it was found from the above experiments that the medium and high doses, i.e. 1.0g and 5.0g, had better effect than the low dose of 0.2g, and the difference between the 1.0g and 5.0g doses was not very large. Therefore, according to "evaluation program and detection method for health food functionality": the result is positive in any two aspects of cellular immunity function, humoral immunity function, macrophage function and NK cell activity, and the tested sample can be judged to have the function of enhancing the immunity, so that the bifidobacterium animalis NSY0201 and the freeze-dried powder thereof can be judged to have the effect of improving the immunity of the animal body.
The above embodiments are only used for illustrating the technical solution of the present invention, and not for limiting the same; although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that modifications may be made to the embodiments described in the foregoing embodiments, or equivalents may be substituted for some of the features thereof; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions.
Sequence listing
<110> Qingdao Norson Biotechnology, Inc
<120> bifidobacterium animalis NSY0201 and application thereof in preparing health care products or medicines for enhancing immunity of organisms
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1462
<212> DNA
<213> Bifidobacterium animalis (Bifidobacterium animalis)
<400> 1
gatgaacgct ggcggcgtgc ttaacacatg caagtcgaac gggatccctg gcagcttgct 60
gtcggggtga gagtggcgaa cgggtgagta atgcgtgacc aacctgccct gtgcaccgga 120
atagctcctg gaaacgggtg gtaataccgg atgctccgct ccatcgcatg gtggggtggg 180
aaatgctttt gcggcatggg atggggtcgc gtcctatcag cttgttggcg gggtgatggc 240
ccaccaaggc gttgacgggt agccggcctg aragggtgac cggccacatt gggactgaga 300
tacggcccag actcctacgg gaggcagcag tggggaatat tgcacaatgg gcgcaagcct 360
gatgcagcga cgccgcgtgc gggatggagg ccttcgggtt gtaaaccgct tttgttcaag 420
ggcaaggcac ggtttcggcc gtgttgagtg gattgttcga ataagcaccg gctaactacg 480
tgccagcagc cgcggtaata cgtagggtgc gagcgttatc cggatttatt gggcgtaaag 540
ggctcgtagg cggttcgtcg cgtccggtgt gaaagtccat cgcctaacgg tggatctgcg 600
ccgggtacgg gcgggctgga gtgcggtagg ggagactgga attcccggtg taacgtgtgg 660
aatgtgtaga tatcgggaag aacaccaatg gcgaaggcag gtctctgggc ccgtcactga 720
cgctgaggag cgaaagcgtg aggagcgaac aggattagat accctggtag tccacgccgt 780
aaacggtgga tgctggatgt ggggcccttt ccacgggtcc cgtgtcggag ccaacgcgtt 840
aagcatcccg cctggggagt acggccgcaa ggctaaaact caaagaaatt gacgggggcc 900
cgcacaagcg gcggagcatg cggattaatt cgatgcaacg cgaagaacct tacctgggct 960
tgacatgtgc cggatcgccg tggagacacg gtttcccttc ggggccggtt cacaggtggt 1020
gcatggtcgt cgtcagctcg tgtcgtgaga tgttgggtta agtcccgcaa cgagcgcaac 1080
cctcgccgca tgttgccagc gggtgatgcc gggaactcat gtgggaccgc cggggtcaac 1140
tcggaggaag gtggggatga cgtcagatca tcatgcccct tacgtccagg gcttcacgca 1200
tgctacaatg gccggtacaa cgcggtgcga cacggtgacg tggggcggat cgctgaaaac 1260
cggtctcagt tcggatcgca gtctgcaact cgactgcgtg aaggcggagt cgctagtaat 1320
cgcggatcag caacgccgcg gtgaatgcgt tcccgggcct tgtacacacc gcccgtcaag 1380
tcatgaaagt gggtagcacc cggagccggt ggcccgaccc ttgtgggggg agccgtctaa 1440
ggtgagactc gtgattggga ct 1462
Claims (4)
1. The bifidobacterium animalis NSY0201 is characterized in that the bifidobacterium animalis NSY0201 is named as bifidobacterium animalisBifidobacterium animalisAnd the product is preserved in China center for type culture Collection with the preservation number of CTCC M2021629.
2. Use of Bifidobacterium animalis NSY0201 in claim 1 for preparing health products or pharmaceuticals for enhancing immunity, wherein the health products or pharmaceuticals comprise viable bacteria in an amount of not less than 4 x 1011 CFU/g animal bifidobacterium NSY0201 freeze-dried powder.
3. The use of claim 2, wherein the bifidobacterium animalis NSY0201 lyophilized powder is in an amount of 0.2% -5% by weight of the health care product or medicament.
4. The use of claim 2, wherein the bifidobacterium animalis NSY0201 freeze-dried powder is prepared by the following steps: activating the bifidobacterium animalis NSY0201, inoculating the bifidobacterium animalis NSY0201 in an improved MRS liquid culture medium in an inoculation amount of 2-5% (v/v), culturing at 37 ℃ for 14-18 h, continuously culturing for 2 generations of expansion culture, centrifuging and cleaning the obtained fermentation liquor at low temperature, uniformly mixing the collected bacterial sludge and a freeze-drying protective agent in a ratio of 1:1.5-2, fully emulsifying, and performing vacuum freeze-drying to obtain the bifidobacterium animalis NSY0201 freeze-dried powder.
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| CN101260377A (en) * | 2008-03-14 | 2008-09-10 | 内蒙古蒙牛乳业（集团）股份有限公司 | Animal bifidobacteria and use thereof |
| KR20170055093A (en) * | 2015-11-10 | 2017-05-19 | 대한민국(농촌진흥청장) | Bifidobacterium animalis with immune enhancing effect |
| CN111110703A (en) * | 2020-01-13 | 2020-05-08 | 华中农业大学 | Bifidobacterium animalis and application of compound bacterium preparation prepared from the same in preparation of medicines for treating or preventing avian influenza virus infection |
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| CN101260377A (en) * | 2008-03-14 | 2008-09-10 | 内蒙古蒙牛乳业（集团）股份有限公司 | Animal bifidobacteria and use thereof |
| KR20170055093A (en) * | 2015-11-10 | 2017-05-19 | 대한민국(농촌진흥청장) | Bifidobacterium animalis with immune enhancing effect |
| CN111110703A (en) * | 2020-01-13 | 2020-05-08 | 华中农业大学 | Bifidobacterium animalis and application of compound bacterium preparation prepared from the same in preparation of medicines for treating or preventing avian influenza virus infection |
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