CN112458015B - Bifidobacterium longum subspecies longum i772, and separation and purification method and application thereof - Google Patents
Bifidobacterium longum subspecies longum i772, and separation and purification method and application thereof Download PDFInfo
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- CN112458015B CN112458015B CN202011361316.5A CN202011361316A CN112458015B CN 112458015 B CN112458015 B CN 112458015B CN 202011361316 A CN202011361316 A CN 202011361316A CN 112458015 B CN112458015 B CN 112458015B
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- 241001608472 Bifidobacterium longum Species 0.000 title claims abstract description 60
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- 238000000034 method Methods 0.000 title abstract description 14
- 238000000746 purification Methods 0.000 title abstract description 12
- 238000000926 separation method Methods 0.000 title abstract description 12
- 210000004185 liver Anatomy 0.000 claims abstract description 20
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- 239000006041 probiotic Substances 0.000 claims abstract description 16
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- 244000005700 microbiome Species 0.000 claims abstract 2
- 238000002156 mixing Methods 0.000 claims description 6
- 108090000623 proteins and genes Proteins 0.000 claims description 5
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- 101150099542 tuf gene Proteins 0.000 claims description 4
- 241000185999 Bifidobacterium longum subsp. longum Species 0.000 claims description 3
- 238000009629 microbiological culture Methods 0.000 claims description 3
- 238000002360 preparation method Methods 0.000 claims description 3
- 241000186016 Bifidobacterium bifidum Species 0.000 claims description 2
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- 150000001875 compounds Chemical class 0.000 claims description 2
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- 238000001514 detection method Methods 0.000 description 3
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- 208000036649 Dysbacteriosis Diseases 0.000 description 1
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- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 206010019728 Hepatitis alcoholic Diseases 0.000 description 1
- 208000017604 Hodgkin disease Diseases 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
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- 206010067125 Liver injury Diseases 0.000 description 1
- BYPFEZZEUUWMEJ-UHFFFAOYSA-N Pentoxifylline Chemical compound O=C1N(CCCCC(=O)C)C(=O)N(C)C2=C1N(C)C=N2 BYPFEZZEUUWMEJ-UHFFFAOYSA-N 0.000 description 1
- 241000555745 Sciuridae Species 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
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- 239000003246 corticosteroid Substances 0.000 description 1
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- -1 diamine citrate Chemical class 0.000 description 1
- 229910000396 dipotassium phosphate Inorganic materials 0.000 description 1
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- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 1
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Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/52—Adding ingredients
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/745—Bifidobacteria
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
- A61P39/02—Antidotes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/02—Separating microorganisms from their culture media
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/51—Bifidobacterium
- A23V2400/533—Longum
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The invention belongs to the technical field of bioengineering, and discloses a bifidobacterium longum subspecies i772, a separation and purification method and application thereof, wherein the strain is preserved in the common microorganism center of China Committee for culture Collection of microorganisms with the preservation number of CGMCC NO. 19856; the Bifidobacterium longum subspecies is obtained by enriching, screening and continuously purifying target strains in feces of breast-fed infants or children. Compared with the existing bifidobacterium longum subspecies longum, the bifidobacterium longum subspecies screened by the invention has the application of regulating the balance of intestinal flora and relieving the damage of alcohol to the liver, and the strain is used for preparing drinks, foods, medicines or probiotic products with the intestinal regulation function.
Description
Technical Field
The invention belongs to the technical field of bioengineering, and relates to a strain, a screening method and application thereof, in particular to bifidobacterium longum subspecies longum i772, and a separation and purification method and application thereof.
Background
Alcohol drinking may lead to Alcoholic Liver Disease (ALD). Ethanol can cause the damage of intestinal barrier and dysbacteriosis of ALD patients, alcohol drinkers without liver diseases, and patients with alcoholic hepatitis and alcoholic cirrhosis with higher endotoxin level in plasma than normal people, thereby triggering organism inflammatory reaction.
Current approaches to treating ALD include mainly alcohol abstinence and nutritional support as well as drug therapy. Corticosteroids, pentoxifylline, anti-tnf α antibodies, antioxidants, etc. have been used clinically, but the efficacy of these drugs is limited.
At present, probiotics such as lactobacillus, bifidobacterium, lactobacillus acidophilus and the like are widely applied to the fields of bioengineering, food safety and life health, so that the regulation and intervention on human bodies by utilizing the probiotics are more and more extensive.
Disclosure of Invention
The invention aims to solve the technical problem of providing a bifidobacterium longum subspecies i772 to achieve the purposes of adjusting the balance of intestinal flora and relieving the damage of alcohol to the liver;
another objective of the present invention is to provide a method for preparing bifidobacterium longum subspecies i772, so as to achieve the purpose of rapidly, effectively and accurately screening the strain;
the third objective of the present invention is to provide the application of the bifidobacterium longum subspecies longum i772 mentioned above.
In order to achieve the purpose, the invention adopts the technical scheme that:
the Bifidobacterium longum subspecies i772, the Bifidobacterium longum subspecies i772 is preserved in the common microorganism center of China general microbiological culture Collection center with the preservation number of CGMCC NO.19856, and the Latin name is Bifidobacterium longum subsp.
The sequence of the 16SrDNA gene of the bifidobacterium longum subspecies longum i772 is defined as follows:
AGTCGAACGGGATCCATCAGGCTTTGCTTGGTGGTGAGAGTGGCGAACGGGTGAGTAATGCGTG ACCGACCTGCCCCATACACCGGAATAGCTCCTGGAAACGGGTGGTAATGCCGGATGCTCCAGTTGATC GCATGGTCTTCTGGGAAAGCTTTCGCGGTATGGGATGGGGTCGCGTCCTATCAGCTTGACGGCGGGGT AACGGCCCACCGTGGCTTCGACGGGTAGCCGGCCTGAGAGGGCGACCGGCCACATTGGGACTGAGATA CGGCCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGCGCAAGCCTGATGCAGCGA CGCCGCGTGAGGGATGGAGGCCTTCGGGTTGTAAACCTCTTTTATCGGGGAGCAAGCGAGAGTGAGTT TACCCGTTGAATAAGCACCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGTGCAAGCGTTA TCCGGAATTATTGGGCGTAAAGGGCTCGTAGGCGGTTCGTCGCGTCCGGTGTGAAAGTCCATCGCTTA ACGGTGGATCCGCGCCGGGTACGGGCGGGCTTGAGTGCGGTAGGGGAGACTGGAATTCCCGGTGTAAC GGTGGAATGTGTAGATATCGGGAAGAACACCAATGGCGAAGGCAGGTCTCTGGGCCGTTACTGACGCT GAGGAGCGAAAGCGTGGGGAGCGAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGGTGGATG CTGGATGTGGGGCCCGTTCCACGGGTTCCGTGTCGGAGCTAACGCGTTAAGCATCCCGCCTGGGGAGT ACGGCCGCAAGGCTAAACTCAAAGAAATTGACGGGGGCCCGCACAAGCGGCGGAGCATGCGGATTAAT TCGATGCAACGCGAAGAACCTTACCTGGGCTTGACATGTTCCCGACGGTCGTAGAGATACGGCTTCCC TTCGGGGCGGGTTCACAGGTGGTGCATGGTCGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCC CGCAACGAGCGCAACCCTCGCCCCGTGTTGCCAGCGGATTATGCCGGGAACTCACGGGGGACCGCCGG GGTTAACTCGGAGGAAGGTGGGGATGACGTCAGATCATCATGCCCCTTACGTCCAGGGCTTCACGCAT GCTACAATGGCCGGTACAACGGGATGCGACGCGGCGACGCGGAGCGGATCCCTGAAAACCGGTCTCAG TTCGGATCGCAGTCTGCAACTCGACTGCGTGAAGGCGGAGTCGCTAGTAATCGCGAATCAGCAACGTC GCGGTGAATGCGTTCCCGGGCCTTGTACACACCGCCCGTCAAGTCATGAAAGTGGGCAGCACCCGAAG CCGGTGGCCTAACCCCTTGTGGGATGGAGCC。
as another limitation of the present invention, the tuf gene sequence of said bifidobacterium longum subspecies longum i772 is as follows:
CCGCGGTATCACCATCAACATCGCCCACATCGAGTACCAGACCGAGAAGCGTCACTACGCTCAC GTCGACTGCCCGGGCCACGCCGACTTCGTGAAGAACATGATTACCGGTGCTGCCCAGATGGATGGCGC TATCCTCGTTGTGGCCGCCACCGACGGCCCGATGGCCCAGACTCGCGAGCACGTGCTGCTCGCCCGTC AGGTTGGCGTTCCGAAGATCCTCGTCGCCCTGAACAAGTGCGACATGGTCGACGATGAAGAGCTCATC GAGCTCGTCGAAGAAGAGGTCCGCGACCTCCTCGACGAGAACGGCTTCGACCGTGACTGCCCGGTCAT CCACACCTCCGCTTACGGTGCTCTGCACGACGACGCTCCGGACCACGAGAAGTGGGTCCAGTCCGTTA AGGACCTCATGGACGCTGTCGACGACTACATCCCGACCCCGGTTCACGACCTGGACAAGCCGTTCCTG ATGCCGATCGAGGACGTCTTCACCATCTCCGGCCGTGGTACCGTTGTCACCGGTCGTGTCGAGCGTGG CCAGCTGGCCGTCAACACCCCGGTCGAGATCGTTGGTATCCGTCCGACCCAGCAGACCACCGTCACCT CCATCGAGACCTTCCACAAGACCATGGACGCCTGCGAGGCTGGCGACAACACCGGTCTGCTTCTGCGT GGTCTCGGCCGTGACGATGTCGAGCGTGGCCAGGTTGTGGCCAAGCCGGGCTCCGTCACCCCGCACAC CAAGTTCGAGGGCGAAGTCTACGTGCTGACCAAGGACGAAGGCGGCCGTCACTCGCCGTTCTTCTCCA ACTACCGTCCGCAGTTCTACTTCCGCACCACCGACGTCACCGGCGTCATCGAGCTGCCGG。
as a third limitation of the present invention, said Bifidobacterium longum subspecies i772 is isolated and screened from the feces of a breast-fed infant or young child.
The invention also provides a separation and purification method of the bifidobacterium longum subspecies longum i772, which comprises the following steps of:
s1, collecting a sample
Adding feces of breast-fed infants or infants into physiological saline, and fully and uniformly mixing to obtain a sample A;
s2, enriching the sample
Adding the sample A into an improved MRS liquid culture medium, and carrying out anaerobic culture for 62-82 h at 35-40 ℃ to obtain a culture solution B; wherein:
the improved MRS liquid culture medium is an MRS liquid culture medium added with 0.5 per mill of cysteine by mass;
the volume ratio of the sample A to the improved MRS liquid culture medium is 1: 10-100;
s3, strain separation and screening
Taking culture solution B, and performing gradient with 10 times of sterile physiological saline with concentration of 0.9%Degree-multiplied dilution, in turn, gradient dilution 10-1、10-2、10-3、10-4、10-5Multiplying to obtain bacterial suspension C1-C5;
taking the improved MRS solid culture medium, melting, pouring the melted improved MRS solid culture medium into the first to fifth culture dishes respectively, and obtaining culture media D1-D5 after cooling and complete solidification; wherein the improved MRS solid culture medium is obtained by adding 15% of agar in parts by mass into each 1000mL of improved MRS liquid culture medium;
respectively sucking 0.1mL of bacterial suspension C1-C5, coating the bacterial suspension on culture media D1-D5 in a one-to-one correspondence manner, inverting the flat plate, placing the flat plate in an environment with the temperature of 35-40 ℃ for anaerobic culture for 62-82 h, and observing the growth condition of bacterial colonies;
after the plate has the typical bacteria, selecting typical single bacterial colony E;
s4, strain purification
Selecting a selected single colony E, streaking and inoculating a single colony E culture on an improved MRS solid culture medium, and culturing for 62-82 h at 35-40 ℃ under an anaerobic environment; continuously culturing for three times; obtaining a pure culture F, namely the bifidobacterium longum subspecies i 772.
As a limitation of the above separation and purification method, the Bifidobacterium longum subspecies longum i772 strain can be preserved as follows: mixing the pure culture F with sterile glycerol with the mass portion of 50% according to the proportion of 1:1, storing in a sterile environment at the temperature of minus 80-minus 70 ℃, and simultaneously inoculating to a test tube slant of a modified MRS solid culture medium for temporary storage.
As another limitation of the above separation and purification method, the modified MRS liquid medium is composed of 10 parts by weight of casein peptone, 10 parts by weight of beef extract, 5 parts by weight of yeast extract, 20 parts by weight of glucose, 5 parts by weight of sodium acetate, 2 parts by weight of diamine citrate, 1 part by weight of Tween-80, 2 parts by weight of K2HPO40.2 part by weight of MgSO4·7H2O, 0.05 part by weight of MnSO4·7H2O, 0.5 part by weight of cysteine and 1000 parts by volume of distilled water; wherein the corresponding proportion relationship between the parts by weight and the parts by volume is g: mL.
The bacteriological properties of Bifidobacterium longum subspecies longum i772 of the present invention are as follows:
(1) strain characteristic experiment
Carrying out anaerobic culture on the separated and purified strain at 35-40 ℃ for 62-82 h, and respectively carrying out observation and experiment on strain characteristics, wherein the results are as follows:
note: "-" indicates no relevant essential feature;
(2) molecular biological identification
The bacterial strain is subjected to molecular biological identification, DNA extraction and PCR amplification, 16Sr RNA and tuf gene sequence sequencing is carried out, and the sequencing result is as follows:
the 16Sr gene sequence of Bifidobacterium longum subspecies i772 is as follows:
AGTCGAACGGGATCCATCAGGCTTTGCTTGGTGGTGAGAGTGGCGAACGGGTGAGTAATGCGTG ACCGACCTGCCCCATACACCGGAATAGCTCCTGGAAACGGGTGGTAATGCCGGATGCTCCAGTTGATC GCATGGTCTTCTGGGAAAGCTTTCGCGGTATGGGATGGGGTCGCGTCCTATCAGCTTGACGGCGGGGT AACGGCCCACCGTGGCTTCGACGGGTAGCCGGCCTGAGAGGGCGACCGGCCACATTGGGACTGAGATA CGGCCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGCGCAAGCCTGATGCAGCGA CGCCGCGTGAGGGATGGAGGCCTTCGGGTTGTAAACCTCTTTTATCGGGGAGCAAGCGAGAGTGAGTT TACCCGTTGAATAAGCACCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGTGCAAGCGTTA TCCGGAATTATTGGGCGTAAAGGGCTCGTAGGCGGTTCGTCGCGTCCGGTGTGAAAGTCCATCGCTTA ACGGTGGATCCGCGCCGGGTACGGGCGGGCTTGAGTGCGGTAGGGGAGACTGGAATTCCCGGTGTAAC GGTGGAATGTGTAGATATCGGGAAGAACACCAATGGCGAAGGCAGGTCTCTGGGCCGTTACTGACGCT GAGGAGCGAAAGCGTGGGGAGCGAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGGTGGATG CTGGATGTGGGGCCCGTTCCACGGGTTCCGTGTCGGAGCTAACGCGTTAAGCATCCCGCCTGGGGAGT ACGGCCGCAAGGCTAAACTCAAAGAAATTGACGGGGGCCCGCACAAGCGGCGGAGCATGCGGATTAAT TCGATGCAACGCGAAGAACCTTACCTGGGCTTGACATGTTCCCGACGGTCGTAGAGATACGGCTTCCC TTCGGGGCGGGTTCACAGGTGGTGCATGGTCGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCC CGCAACGAGCGCAACCCTCGCCCCGTGTTGCCAGCGGATTATGCCGGGAACTCACGGGGGACCGCCGG GGTTAACTCGGAGGAAGGTGGGGATGACGTCAGATCATCATGCCCCTTACGTCCAGGGCTTCACGCAT GCTACAATGGCCGGTACAACGGGATGCGACGCGGCGACGCGGAGCGGATCCCTGAAAACCGGTCTCAG TTCGGATCGCAGTCTGCAACTCGACTGCGTGAAGGCGGAGTCGCTAGTAATCGCGAATCAGCAACGTC GCGGTGAATGCGTTCCCGGGCCTTGTACACACCGCCCGTCAAGTCATGAAAGTGGGCAGCACCCGAAG CCGGTGGCCTAACCCCTTGTGGGATGGAGCC。
the thf gene sequence of bifidobacterium longum subspecies longum i772 is as follows:
CCGCGGTATCACCATCAACATCGCCCACATCGAGTACCAGACCGAGAAGCGTCACTACGCTCAC GTCGACTGCCCGGGCCACGCCGACTTCGTGAAGAACATGATTACCGGTGCTGCCCAGATGGATGGCGC TATCCTCGTTGTGGCCGCCACCGACGGCCCGATGGCCCAGACTCGCGAGCACGTGCTGCTCGCCCGTC AGGTTGGCGTTCCGAAGATCCTCGTCGCCCTGAACAAGTGCGACATGGTCGACGATGAAGAGCTCATC GAGCTCGTCGAAGAAGAGGTCCGCGACCTCCTCGACGAGAACGGCTTCGACCGTGACTGCCCGGTCAT CCACACCTCCGCTTACGGTGCTCTGCACGACGACGCTCCGGACCACGAGAAGTGGGTCCAGTCCGTTA AGGACCTCATGGACGCTGTCGACGACTACATCCCGACCCCGGTTCACGACCTGGACAAGCCGTTCCTG ATGCCGATCGAGGACGTCTTCACCATCTCCGGCCGTGGTACCGTTGTCACCGGTCGTGTCGAGCGTGG CCAGCTGGCCGTCAACACCCCGGTCGAGATCGTTGGTATCCGTCCGACCCAGCAGACCACCGTCACCT CCATCGAGACCTTCCACAAGACCATGGACGCCTGCGAGGCTGGCGACAACACCGGTCTGCTTCTGCGT GGTCTCGGCCGTGACGATGTCGAGCGTGGCCAGGTTGTGGCCAAGCCGGGCTCCGTCACCCCGCACAC CAAGTTCGAGGGCGAAGTCTACGTGCTGACCAAGGACGAAGGCGGCCGTCACTCGCCGTTCTTCTCCA ACTACCGTCCGCAGTTCTACTTCCGCACCACCGACGTCACCGGCGTCATCGAGCTGCCGG；
the gene sequence alignment was finally determined to be bifidobacterium longum subspecies longum i 772.
The invention also provides an application of the bifidobacterium longum subspecies i772, and the bifidobacterium longum subspecies i772 is used for regulating the balance of intestinal flora and relieving the damage of alcohol to the liver.
As a limitation of the application of the Bifidobacterium longum subspecies i772, the Bifidobacterium longum subspecies i772 strain can be used for preparing drinks, foods, medicines or probiotic products with intestinal tract regulation function.
As a further limitation of the use of the bifidobacterium longum subspecies longum i772 mentioned above, the probiotic preparation prepared from said strain of bifidobacterium longum subspecies longum i772 is a probiotic comprising only said strain of bifidobacterium longum subspecies longum i 772; or the compound probiotics is prepared by mixing the bifidobacterium longum subspecies longum i772 with lactobacillus paracasei N1115, lactobacillus rhamnosus X253 and animal bifidobacterium bifidum subspecies i 797.
Due to the adoption of the technical scheme, compared with the prior art, the invention has the technical progress that:
bifidobacterium longum subspecies longum i772 can effectively relieve the damage of alcohol to the liver. Alanine aminotransferase (also called glutamic-pyruvic transaminase, ALT) is mainly distributed in liver cytoplasm, aspartate aminotransferase (also called glutamic-oxaloacetic transaminase, AST) is mainly distributed in liver cytoplasm and liver cell mitochondria, gamma-glutamyl transpeptidase (GGT) is mainly distributed in bile capillary and liver cell microsomes, and when pathogenic factors (such as alcohol) cause liver cell degeneration and increase of cell membrane permeability, ALT, AST and GGT are released, so that the content of the three in serum is remarkably increased. After the strain is used, the reduction of ALT, AST and GGT contents in the liver of a patient can be detected, and the strain can be proved to have the function of relieving the damage degree of alcohol to the liver;
the bacterium also has good intestinal tract regulating effect;
the bifidobacterium longum subspecies longum i772 provided by the invention has wide application range, not only can be used for preparing probiotic products, but also can be used for preparing drinks, foods or medicines.
The present invention is further illustrated by the following specific examples, which are to be construed as merely illustrative, and not limitative of the remainder of the disclosure.
Example 1A Bifidobacterium longum subspecies longum i772 and a method for separating and purifying the same
First, strain information
Bifidobacterium longum subsp.772 is separated and screened from feces of breast-fed infants or children, which has been preserved in China general microbiological culture Collection center at 20.5.2020, with the address of No. 3 of Beijing Hodgkin's area, West Lu No.1, CGMCC No.19856, and the Latin name of Bifidobacterium longum subsp.
Separation and purification method of Bifidobacterium longum subspecies i772
The separation and purification method is sequentially carried out according to the following steps:
s1, collecting a sample
Taking 20g of feces of a breast-fed infant or a baby by using a sterile sampling spoon, adding the feces into 100mL of normal saline, and fully and uniformly mixing to obtain a sample A;
s2, enriching the sample
Adding 2mL of sample A into 100mL of improved MRS liquid culture medium, and carrying out anaerobic culture at 35-40 ℃ for 62-82 h to obtain a culture solution B;
the improved MRS liquid culture medium is an MRS liquid culture medium added with 0.5 per mill of cysteine by mass;
the volume ratio of the sample A to the improved MRS liquid culture medium is 1: 10-100;
s3, strain separation and screening
Taking 1mL of culture solution B, diluting with sterile physiological saline with concentration of 0.9% by 10 times of gradient multiplication, sequentially diluting with gradient 10-1、10-2、10-3、10-4、10-5Multiplying to obtain bacterial suspension C1-C5;
taking an improved MRS solid culture medium, melting, respectively pouring into first to fifth culture dishes, cooling and completely solidifying to obtain culture media D1 to D5, respectively sucking 0.1mL of bacterial suspension C1 to C5, respectively, coating the bacterial suspension C1 to C5 on the culture media D1 to D5 in a one-to-one correspondence manner, inverting the flat plate, placing the flat plate in an environment at 35 to 40 ℃ for anaerobic culture for 62 to 82 hours, and observing the growth condition of bacterial colonies;
the improved MRS solid culture medium is a solid culture medium obtained by adding 15 mass percent of agar into every 1000mL of improved MRS liquid culture medium;
after the plate has the typical bacteria, selecting typical single bacterial colony E;
s4, strain purification
Selecting a selected single colony E, streaking and inoculating a single colony E culture on an improved MRS solid culture medium, and culturing for 62-82 h in an anaerobic environment at the temperature of 35-40 ℃; continuously culturing for three times; obtaining a pure culture F, namely the bifidobacterium longum subspecies i 772.
S5, preservation of strains
Mixing the pure culture F with sterile glycerol with the mass portion of 50% according to the proportion of 1:1, storing in a sterile environment at the temperature of minus 80-minus 70 ℃, and simultaneously inoculating to a test tube slant of a modified MRS solid culture medium for temporary storage.
Example 2 bacteriological characteristics of the species Bifidobacterium longum subspecies longum i772
Strain characteristic experiment
Carrying out anaerobic culture on the separated and purified strain at 35-40 ℃ for 62-82 h, and respectively carrying out observation and experiment on strain characteristics, wherein the results are shown in table 1:
TABLE 1 basic characteristics of Bifidobacterium longum subspecies longum i772
Note: "-" indicates no relevant essential feature.
Molecular biology identification
The strain is subjected to molecular biological identification, and 16Sr RNA of bifidobacterium longum subspecies i772 is sequenced through DNA extraction and PCR amplification, and 16Sr RNA is sequenced through tuf gene sequences. Wherein:
the 16SrDNA gene sequence of Bifidobacterium longum subspecies i772 is as follows:
AGTCGAACGGGATCCATCAGGCTTTGCTTGGTGGTGAGAGTGGCGAACGGGTGAGTAATGCGTG ACCGACCTGCCCCATACACCGGAATAGCTCCTGGAAACGGGTGGTAATGCCGGATGCTCCAGTTGATC GCATGGTCTTCTGGGAAAGCTTTCGCGGTATGGGATGGGGTCGCGTCCTATCAGCTTGACGGCGGGGT AACGGCCCACCGTGGCTTCGACGGGTAGCCGGCCTGAGAGGGCGACCGGCCACATTGGGACTGAGATA CGGCCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGCGCAAGCCTGATGCAGCGA CGCCGCGTGAGGGATGGAGGCCTTCGGGTTGTAAACCTCTTTTATCGGGGAGCAAGCGAGAGTGAGTT TACCCGTTGAATAAGCACCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGTGCAAGCGTTA TCCGGAATTATTGGGCGTAAAGGGCTCGTAGGCGGTTCGTCGCGTCCGGTGTGAAAGTCCATCGCTTA ACGGTGGATCCGCGCCGGGTACGGGCGGGCTTGAGTGCGGTAGGGGAGACTGGAATTCCCGGTGTAAC GGTGGAATGTGTAGATATCGGGAAGAACACCAATGGCGAAGGCAGGTCTCTGGGCCGTTACTGACGCT GAGGAGCGAAAGCGTGGGGAGCGAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGGTGGATG CTGGATGTGGGGCCCGTTCCACGGGTTCCGTGTCGGAGCTAACGCGTTAAGCATCCCGCCTGGGGAGT ACGGCCGCAAGGCTAAACTCAAAGAAATTGACGGGGGCCCGCACAAGCGGCGGAGCATGCGGATTAAT TCGATGCAACGCGAAGAACCTTACCTGGGCTTGACATGTTCCCGACGGTCGTAGAGATACGGCTTCCC TTCGGGGCGGGTTCACAGGTGGTGCATGGTCGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCC CGCAACGAGCGCAACCCTCGCCCCGTGTTGCCAGCGGATTATGCCGGGAACTCACGGGGGACCGCCGG GGTTAACTCGGAGGAAGGTGGGGATGACGTCAGATCATCATGCCCCTTACGTCCAGGGCTTCACGCAT GCTACAATGGCCGGTACAACGGGATGCGACGCGGCGACGCGGAGCGGATCCCTGAAAACCGGTCTCAG TTCGGATCGCAGTCTGCAACTCGACTGCGTGAAGGCGGAGTCGCTAGTAATCGCGAATCAGCAACGTC GCGGTGAATGCGTTCCCGGGCCTTGTACACACCGCCCGTCAAGTCATGAAAGTGGGCAGCACCCGAAG CCGGTGGCCTAACCCCTTGTGGGATGGAGCC；
the thf gene sequence of bifidobacterium longum subspecies longum i772 is as follows:
CCGCGGTATCACCATCAACATCGCCCACATCGAGTACCAGACCGAGAAGCGTCACTACGCTCAC GTCGACTGCCCGGGCCACGCCGACTTCGTGAAGAACATGATTACCGGTGCTGCCCAGATGGATGGCGC TATCCTCGTTGTGGCCGCCACCGACGGCCCGATGGCCCAGACTCGCGAGCACGTGCTGCTCGCCCGTC AGGTTGGCGTTCCGAAGATCCTCGTCGCCCTGAACAAGTGCGACATGGTCGACGATGAAGAGCTCATC GAGCTCGTCGAAGAAGAGGTCCGCGACCTCCTCGACGAGAACGGCTTCGACCGTGACTGCCCGGTCAT CCACACCTCCGCTTACGGTGCTCTGCACGACGACGCTCCGGACCACGAGAAGTGGGTCCAGTCCGTTA AGGACCTCATGGACGCTGTCGACGACTACATCCCGACCCCGGTTCACGACCTGGACAAGCCGTTCCTG ATGCCGATCGAGGACGTCTTCACCATCTCCGGCCGTGGTACCGTTGTCACCGGTCGTGTCGAGCGTGG CCAGCTGGCCGTCAACACCCCGGTCGAGATCGTTGGTATCCGTCCGACCCAGCAGACCACCGTCACCT CCATCGAGACCTTCCACAAGACCATGGACGCCTGCGAGGCTGGCGACAACACCGGTCTGCTTCTGCGT GGTCTCGGCCGTGACGATGTCGAGCGTGGCCAGGTTGTGGCCAAGCCGGGCTCCGTCACCCCGCACAC CAAGTTCGAGGGCGAAGTCTACGTGCTGACCAAGGACGAAGGCGGCCGTCACTCGCCGTTCTTCTCCA ACTACCGTCCGCAGTTCTACTTCCGCACCACCGACGTCACCGGCGTCATCGAGCTGCCGG。
example 3 experiment for alleviating alcoholic liver injury using Bifidobacterium longum subspecies longum i772
Experimental animal and material
Experimental animals including 80 clean male SD rats with body weight of 130 + -10 g
Experimental materials: basal feed (Experimental animal center of Hebei medical university), experimental padding and squirrel cage stored at normal temperature
Experimental strains: bifidobacterium longum subspecies longum i772
Establishing and grouping animal models
All rats are raised in cages in the same room, 5 rats are raised in each cage, the rats freely eat and drink water, and the day and night illumination change period is 12 hours, and the rats are dark for 12 hours. The environmental temperature is controlled at 23 +/-2 ℃, and the humidity is controlled at 55-65%. The experiment was started 1 week after the pilot animals were purchased and acclimatized in the animal feeding facility.
40 male SD rats were randomly divided into a normal control group (n-8) and a pre-treated group of alcohol extracts (n-32). The alcohol dried pre-groups were divided into group A (blank group), group B (low dose treatment group), group C (medium dose treatment group), and group D (high dose treatment group), with 8 per group. Normal diet of control group; the drinking water was replaced with 20% volume fraction of food grade ethanol in the alcohol drying group.
In addition, the number of live bacteria of the group B which are fed with 10mg/Kg is 3 multiplied by 10 according to the re-irrigation of the mice each day10CFU/mL of Bifidobacterium longum subspecies i772 probiotic powder aqueous solution.
The number of live bacteria of the group C is 3 multiplied by 10 according to the re-irrigation of the mice per day and the number of the live bacteria is 50mg/Kg10CFU/mL of Bifidobacterium longum subspecies i772 probiotic powder aqueous solution.
D group re-feeds 100mg/Kg live bacteria 3X 10 per day according to mouse body10CFU/mL of Bifidobacterium longum subspecies i772 probiotic powder aqueous solution.
Feeding for 6 weeks, fasting for 12 hr, collecting blood from eyeball, standing at room temperature for solidification, centrifuging at 3000r for 15min, collecting serum, and storing at-80 deg.C. After blood collection, the liver was rapidly discarded, the wet weight of the liver was weighed, and the liver coefficient was calculated (liver coefficient ═ liver weight/body weight × 100%).
The right lobe of mouse liver is fixed in 10% neutral formalin, the material is obtained conventionally, dehydrated, embedded in paraffin, sliced (4 μm thick), and stained by HE, and pathological professional is used to examine liver lesion by reading the slice under optical microscope.
The detection kit is used for detecting alanine aminotransferase (also known as glutamic-pyruvic transaminase, ALT), aspartate aminotransferase (also known as glutamic-oxaloacetic transaminase, AST) and gamma-glutamyl transpeptidase (GGT).
And (3) detection results:
note: represents significant difference compared to group a (p < 0.05); represents a very significant difference compared to group a (p <0.01)
Results and analysis
ALT is distributed mainly in the hepatocyte cytoplasm, AST is distributed mainly in the hepatocyte cytoplasm and in the hepatocyte mitochondria ("machinery" responsible for providing cellular power within hepatocytes). When the pathogenic factors cause the degeneration of liver cells and the increase of the permeability of cell membranes, ALT is mainly released from the cells; when the liver cells are seriously damaged and necrosed, the AST in mitochondria is released, and the AST in serum is obviously increased. GGT in the liver is mainly localized in the capillary bile duct and in the hepatocyte microsomes, it can reflect hepatocyte necrosis and damage, and can be used for diagnosing and observing the process of alcoholic liver damage of internal hepatic drainage disorder, hepatic obstruction, liver cirrhosis, etc.
Through detection, the high-dose application of the bifidobacterium longum subspecies i772 can achieve the purpose of obviously reducing the liver coefficient of a mouse; improving liver case characteristics; reducing ALT, AST and GGT liver lesion index.
Example 4 application of Bifidobacterium longum subspecies longum i772
The bifidobacterium longum subspecies longum i772 can be used for preparing drinks, foods, medicines or probiotic products with intestinal tract regulating function, and the preparation methods are all common technical means in the field.
Sequence listing
<110> Shijiazhuang Junle Baoru Co Ltd
<120> Bifidobacterium longum subspecies longum i772, and separation and purification method and application thereof
<130> 2020.11.27
<160> 2
<170> PatentIn version 3.5
<210> 1
<211> 1387
<212> DNA
<213> Bifidobacterium longum subsp.longum
<400> 1
agtcgaacgg gatccatcag gctttgcttg gtggtgagag tggcgaacgg gtgagtaatg 60
cgtgaccgac ctgccccata caccggaata gctcctggaa acgggtggta atgccggatg 120
ctccagttga tcgcatggtc ttctgggaaa gctttcgcgg tatgggatgg ggtcgcgtcc 180
tatcagcttg acggcggggt aacggcccac cgtggcttcg acgggtagcc ggcctgagag 240
ggcgaccggc cacattggga ctgagatacg gcccagactc ctacgggagg cagcagtggg 300
gaatattgca caatgggcgc aagcctgatg cagcgacgcc gcgtgaggga tggaggcctt 360
cgggttgtaa acctctttta tcggggagca agcgagagtg agtttacccg ttgaataagc 420
accggctaac tacgtgccag cagccgcggt aatacgtagg gtgcaagcgt tatccggaat 480
tattgggcgt aaagggctcg taggcggttc gtcgcgtccg gtgtgaaagt ccatcgctta 540
acggtggatc cgcgccgggt acgggcgggc ttgagtgcgg taggggagac tggaattccc 600
ggtgtaacgg tggaatgtgt agatatcggg aagaacacca atggcgaagg caggtctctg 660
ggccgttact gacgctgagg agcgaaagcg tggggagcga acaggattag ataccctggt 720
agtccacgcc gtaaacggtg gatgctggat gtggggcccg ttccacgggt tccgtgtcgg 780
agctaacgcg ttaagcatcc cgcctgggga gtacggccgc aaggctaaac tcaaagaaat 840
tgacgggggc ccgcacaagc ggcggagcat gcggattaat tcgatgcaac gcgaagaacc 900
ttacctgggc ttgacatgtt cccgacggtc gtagagatac ggcttccctt cggggcgggt 960
tcacaggtgg tgcatggtcg tcgtcagctc gtgtcgtgag atgttgggtt aagtcccgca 1020
acgagcgcaa ccctcgcccc gtgttgccag cggattatgc cgggaactca cgggggaccg 1080
ccggggttaa ctcggaggaa ggtggggatg acgtcagatc atcatgcccc ttacgtccag 1140
ggcttcacgc atgctacaat ggccggtaca acgggatgcg acgcggcgac gcggagcgga 1200
tccctgaaaa ccggtctcag ttcggatcgc agtctgcaac tcgactgcgt gaaggcggag 1260
tcgctagtaa tcgcgaatca gcaacgtcgc ggtgaatgcg ttcccgggcc ttgtacacac 1320
cgcccgtcaa gtcatgaaag tgggcagcac ccgaagccgg tggcctaacc ccttgtggga 1380
tggagcc 1387
<210> 2
<211> 872
<212> DNA
<213> Bifidobacterium longum subsp.longum
<400> 2
ccgcggtatc accatcaaca tcgcccacat cgagtaccag accgagaagc gtcactacgc 60
tcacgtcgac tgcccgggcc acgccgactt cgtgaagaac atgattaccg gtgctgccca 120
gatggatggc gctatcctcg ttgtggccgc caccgacggc ccgatggccc agactcgcga 180
gcacgtgctg ctcgcccgtc aggttggcgt tccgaagatc ctcgtcgccc tgaacaagtg 240
cgacatggtc gacgatgaag agctcatcga gctcgtcgaa gaagaggtcc gcgacctcct 300
cgacgagaac ggcttcgacc gtgactgccc ggtcatccac acctccgctt acggtgctct 360
gcacgacgac gctccggacc acgagaagtg ggtccagtcc gttaaggacc tcatggacgc 420
tgtcgacgac tacatcccga ccccggttca cgacctggac aagccgttcc tgatgccgat 480
cgaggacgtc ttcaccatct ccggccgtgg taccgttgtc accggtcgtg tcgagcgtgg 540
ccagctggcc gtcaacaccc cggtcgagat cgttggtatc cgtccgaccc agcagaccac 600
cgtcacctcc atcgagacct tccacaagac catggacgcc tgcgaggctg gcgacaacac 660
cggtctgctt ctgcgtggtc tcggccgtga cgatgtcgag cgtggccagg ttgtggccaa 720
gccgggctcc gtcaccccgc acaccaagtt cgagggcgaa gtctacgtgc tgaccaagga 780
cgaaggcggc cgtcactcgc cgttcttctc caactaccgt ccgcagttct acttccgcac 840
caccgacgtc accggcgtca tcgagctgcc gg 872
Claims (6)
1. The Bifidobacterium longum subspecies i772 is characterized in that the Bifidobacterium longum subspecies i772 is preserved in China general microbiological culture Collection center of the Committee for culture Collection of microorganisms with the preservation number of CGMCCNO. 19856.
2. A Bifidobacterium longum subspecies longum i772 of claim 1, wherein the 16SrDNA gene sequence is as follows: AGTCGAACGGGATCCATCAGGCTTTGCTTGGTGGTGAGAGTGGCGAACGGGTGAGTAATGCGTGACCGACCTGCCCCATACACCGGAATAGCTCCTGGAAACGGGTGGTAATGCCGGATGCTCCAGTTGATCGCATGGTCTTCTGGGAAAGCTTTCGCGGTATGGGATGGGGTCGCGTCCTATCAGCTTGACGGCGGGGTAACGGCCCACCGTGGCTTCGACGGGTAGCCGGCCTGAGAGGGCGACCGGCCACATTGGGACTGAGATACGGCCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGCGCAAGCCTGATGCAGCGACGCCGCGTGAGGGATGGAGGCCTTCGGGTTGTAAACCTCTTTTATCGGGGAGCAAGCGAGAGTGAGTTTACCCGTTGAATAAGCACCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGTGCAAGCGTTATCCGGAATTATTGGGCGTAAAGGGCTCGTAGGCGGTTCGTCGCGTCCGGTGTGAAAGTCCATCGCTTAACGGTGGATCCGCGCCGGGTACGGGCGGGCTTGAGTGCGGTAGGGGAGACTGGAATTCCCGGTGTAACGGTGGAATGTGTAGATATCGGGAAGAACACCAATGGCGAAGGCAGGTCTCTGGGCCGTTACTGACGCTGAGGAGCGAAAGCGTGGGGAGCGAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGGTGGATGCTGGATGTGGGGCCCGTTCCACGGGTTCCGTGTCGGAGCTAACGCGTTAAGCATCCCGCCTGGGGAGTACGGCCGCAAGGCTAAACTCAAAGAAATTGACGGGGGCCCGCACAAGCGGCGGAGCATGCGGATTAATTCGATGCAACGCGAAGAACCTTACCTGGGCTTGACATGTTCCCGACGGTCGTAGAGATACGGCTTCCCTTCGGGGCGGGTTCACAGGTGGTGCATGGTCGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTCGCCCCGTGTTGCCAGCGGATTATGCCGGGAACTCACGGGGGACCGCCGGGGTTAACTCGGAGGAAGGTGGGGATGACGTCAGATCATCATGCCCCTTACGTCCAGGGCTTCACGCATGCTACAATGGCCGGTACAACGGGATGCGACGCGGCGACGCGGAGCGGATCCCTGAAAACCGGTCTCAGTTCGGATCGCAGTCTGCAACTCGACTGCGTGAAGGCGGAGTCGCTAGTAATCGCGAATCAGCAACGTCGCGGTGAATGCGTTCCCGGGCCTTGTACACACCGCCCGTCAAGTCATGAAAGTGGGCAGCACCCGAAGCCGGTGGCCTAACCCCTTGTGGGATGGAGCC are provided.
3. Bifidobacterium longum subspecies longum i772 of claim 1, wherein: the tuf gene sequence is as follows: CCGCGGTATCACCATCAACATCGCCCACATCGAGTACCAGACCGAGAAGCGTCACTACGCTCACGTCGACTGCCCGGGCCACGCCGACTTCGTGAAGAACATGATTACCGGTGCTGCCCAGATGGATGGCGCTATCCTCGTTGTGGCCGCCACCGACGGCCCGATGGCCCAGACTCGCGAGCACGTGCTGCTCGCCCGTCAGGTTGGCGTTCCGAAGATCCTCGTCGCCCTGAACAAGTGCGACATGGTCGACGATGAAGAGCTCATCGAGCTCGTCGAAGAAGAGGTCCGCGACCTCCTCGACGAGAACGGCTTCGACCGTGACTGCCCGGTCATCCACACCTCCGCTTACGGTGCTCTGCACGACGACGCTCCGGACCACGAGAAGTGGGTCCAGTCCGTTAAGGACCTCATGGACGCTGTCGACGACTACATCCCGACCCCGGTTCACGACCTGGACAAGCCGTTCCTGATGCCGATCGAGGACGTCTTCACCATCTCCGGCCGTGGTACCGTTGTCACCGGTCGTGTCGAGCGTGGCCAGCTGGCCGTCAACACCCCGGTCGAGATCGTTGGTATCCGTCCGACCCAGCAGACCACCGTCACCTCCATCGAGACCTTCCACAAGACCATGGACGCCTGCGAGGCTGGCGACAACACCGGTCTGCTTCTGCGTGGTCTCGGCCGTGACGATGTCGAGCGTGGCCAGGTTGTGGCCAAGCCGGGCTCCGTCACCCCGCACACCAAGTTCGAGGGCGAAGTCTACGTGCTGACCAAGGACGAAGGCGGCCGTCACTCGCCGTTCTTCTCCAACTACCGTCCGCAGTTCTAC TTCCGCACCACCGACGTCACCGGCGTCATCGAGCTGCCGG are provided.
4. Bifidobacterium longum subspecies longum i772 according to any of the claims 1-3, characterized in that: it is separated and screened from the excrement of breast-fed infants or young children.
5. Use of bifidobacterium longum subsp.longum i772 according to any one of claims 1-4, wherein: the strain is used for preparing beverages, foods, medicines or probiotic products which have the intestinal tract regulating function and can relieve the damage of alcohol to the liver.
6. Use of Bifidobacterium longum subspecies longum i772 according to claim 5, characterized in that: the probiotic preparation is a probiotic comprising only said bifidobacterium longum subspecies longum i 772; or the compound probiotics is prepared by mixing the bifidobacterium longum subspecies longum i772 with lactobacillus paracasei N1115, lactobacillus rhamnosus X253 and animal bifidobacterium bifidum subspecies i 797.
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| CN114568703A (en) * | 2022-02-24 | 2022-06-03 | 重庆市天友乳业股份有限公司 | New application of bifidobacterium lactis |
| CN116555076B (en) * | 2023-03-13 | 2023-11-28 | 广东悦创生物科技有限公司 | Bifidobacterium longum subspecies longum MY1 and application thereof in preparation of food and medicine for relaxing bowels and protecting intestines |
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