CN106167778B - Clostridium tender and culture method and application thereof - Google Patents
Clostridium tender and culture method and application thereof Download PDFInfo
- Publication number
- CN106167778B CN106167778B CN201610698863.XA CN201610698863A CN106167778B CN 106167778 B CN106167778 B CN 106167778B CN 201610698863 A CN201610698863 A CN 201610698863A CN 106167778 B CN106167778 B CN 106167778B
- Authority
- CN
- China
- Prior art keywords
- clostridium leptum
- clostridium
- rcm
- leptum
- culture
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 241000193403 Clostridium Species 0.000 title abstract description 13
- 238000012136 culture method Methods 0.000 title abstract 4
- 241000186569 [Clostridium] leptum Species 0.000 claims abstract description 85
- 239000001257 hydrogen Substances 0.000 claims abstract description 9
- 229910052739 hydrogen Inorganic materials 0.000 claims abstract description 9
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 claims abstract description 8
- 235000013325 dietary fiber Nutrition 0.000 claims abstract description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 21
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 20
- 239000002609 medium Substances 0.000 claims description 17
- 239000012153 distilled water Substances 0.000 claims description 16
- 239000012530 fluid Substances 0.000 claims description 15
- 235000015097 nutrients Nutrition 0.000 claims description 15
- 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 claims description 13
- 239000008103 glucose Substances 0.000 claims description 13
- 238000000034 method Methods 0.000 claims description 13
- 239000000284 extract Substances 0.000 claims description 12
- PLXBWHJQWKZRKG-UHFFFAOYSA-N Resazurin Chemical compound C1=CC(=O)C=C2OC3=CC(O)=CC=C3[N+]([O-])=C21 PLXBWHJQWKZRKG-UHFFFAOYSA-N 0.000 claims description 11
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 claims description 11
- 229920002472 Starch Polymers 0.000 claims description 11
- VLSOAXRVHARBEQ-UHFFFAOYSA-N [4-fluoro-2-(hydroxymethyl)phenyl]methanol Chemical compound OCC1=CC=C(F)C=C1CO VLSOAXRVHARBEQ-UHFFFAOYSA-N 0.000 claims description 11
- 235000015278 beef Nutrition 0.000 claims description 11
- 239000001632 sodium acetate Substances 0.000 claims description 11
- 235000017281 sodium acetate Nutrition 0.000 claims description 11
- 235000019698 starch Nutrition 0.000 claims description 11
- 239000008107 starch Substances 0.000 claims description 11
- 239000012137 tryptone Substances 0.000 claims description 11
- 239000011780 sodium chloride Substances 0.000 claims description 10
- 239000001963 growth medium Substances 0.000 claims description 9
- 238000009630 liquid culture Methods 0.000 claims description 7
- 239000001913 cellulose Substances 0.000 claims description 5
- 229920002678 cellulose Polymers 0.000 claims description 5
- 239000007789 gas Substances 0.000 claims description 5
- 239000007787 solid Substances 0.000 claims description 5
- 239000000758 substrate Substances 0.000 claims description 4
- 229920001817 Agar Polymers 0.000 claims description 3
- 239000008272 agar Substances 0.000 claims description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims description 3
- 235000013406 prebiotics Nutrition 0.000 claims description 3
- 201000010099 disease Diseases 0.000 claims description 2
- 230000000968 intestinal effect Effects 0.000 claims description 2
- 208000023275 Autoimmune disease Diseases 0.000 claims 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims 1
- 239000006071 cream Substances 0.000 claims 1
- 108090000790 Enzymes Proteins 0.000 abstract description 9
- 102000004190 Enzymes Human genes 0.000 abstract description 9
- FERIUCNNQQJTOY-UHFFFAOYSA-N Butyric acid Natural products CCCC(O)=O FERIUCNNQQJTOY-UHFFFAOYSA-N 0.000 abstract description 6
- FERIUCNNQQJTOY-UHFFFAOYSA-M Butyrate Chemical compound CCCC([O-])=O FERIUCNNQQJTOY-UHFFFAOYSA-M 0.000 abstract description 4
- 230000000694 effects Effects 0.000 abstract description 3
- 241000087704 Coelosia tenella Species 0.000 abstract 2
- 239000000203 mixture Substances 0.000 description 18
- 210000004027 cell Anatomy 0.000 description 15
- 238000002474 experimental method Methods 0.000 description 11
- 229940041514 candida albicans extract Drugs 0.000 description 10
- 239000012138 yeast extract Substances 0.000 description 10
- 241000894006 Bacteria Species 0.000 description 9
- 239000008246 gaseous mixture Substances 0.000 description 9
- 238000002347 injection Methods 0.000 description 9
- 239000007924 injection Substances 0.000 description 9
- 229940088598 enzyme Drugs 0.000 description 8
- 238000004321 preservation Methods 0.000 description 8
- 239000000243 solution Substances 0.000 description 8
- 108020004465 16S ribosomal RNA Proteins 0.000 description 6
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 6
- 108090000623 proteins and genes Proteins 0.000 description 6
- 238000009835 boiling Methods 0.000 description 5
- 230000012010 growth Effects 0.000 description 5
- 238000004587 chromatography analysis Methods 0.000 description 4
- 238000001816 cooling Methods 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 239000002808 molecular sieve Substances 0.000 description 4
- 230000000877 morphologic effect Effects 0.000 description 4
- 238000012163 sequencing technique Methods 0.000 description 4
- URGAHOPLAPQHLN-UHFFFAOYSA-N sodium aluminosilicate Chemical compound [Na+].[Al+3].[O-][Si]([O-])=O.[O-][Si]([O-])=O URGAHOPLAPQHLN-UHFFFAOYSA-N 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 108010010803 Gelatin Proteins 0.000 description 3
- 238000012408 PCR amplification Methods 0.000 description 3
- 230000009604 anaerobic growth Effects 0.000 description 3
- 150000001875 compounds Chemical class 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 239000008273 gelatin Substances 0.000 description 3
- 229920000159 gelatin Polymers 0.000 description 3
- 235000019322 gelatine Nutrition 0.000 description 3
- 235000011852 gelatine desserts Nutrition 0.000 description 3
- 229960003444 immunosuppressant agent Drugs 0.000 description 3
- 230000001861 immunosuppressant effect Effects 0.000 description 3
- 239000003018 immunosuppressive agent Substances 0.000 description 3
- 238000005259 measurement Methods 0.000 description 3
- 238000002156 mixing Methods 0.000 description 3
- 150000002772 monosaccharides Chemical class 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 150000004666 short chain fatty acids Chemical class 0.000 description 3
- 235000021391 short chain fatty acids Nutrition 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 229920002488 Hemicellulose Polymers 0.000 description 2
- 206010061218 Inflammation Diseases 0.000 description 2
- 108010009736 Protein Hydrolysates Proteins 0.000 description 2
- 208000037883 airway inflammation Diseases 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 239000000084 colloidal system Substances 0.000 description 2
- 201000006549 dyspepsia Diseases 0.000 description 2
- 210000002919 epithelial cell Anatomy 0.000 description 2
- 239000000835 fiber Substances 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- 150000004676 glycans Chemical class 0.000 description 2
- 230000036737 immune function Effects 0.000 description 2
- 230000004054 inflammatory process Effects 0.000 description 2
- 230000002503 metabolic effect Effects 0.000 description 2
- 239000002773 nucleotide Substances 0.000 description 2
- 125000003729 nucleotide group Chemical group 0.000 description 2
- 229920001542 oligosaccharide Polymers 0.000 description 2
- 150000002482 oligosaccharides Chemical class 0.000 description 2
- 229920001282 polysaccharide Polymers 0.000 description 2
- 239000005017 polysaccharide Substances 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 239000006041 probiotic Substances 0.000 description 2
- 235000018291 probiotics Nutrition 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 238000010186 staining Methods 0.000 description 2
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 description 1
- 229920002134 Carboxymethyl cellulose Polymers 0.000 description 1
- 108010059892 Cellulase Proteins 0.000 description 1
- KZBUYRJDOAKODT-UHFFFAOYSA-N Chlorine Chemical compound ClCl KZBUYRJDOAKODT-UHFFFAOYSA-N 0.000 description 1
- 206010009944 Colon cancer Diseases 0.000 description 1
- 208000011231 Crohn disease Diseases 0.000 description 1
- 108010067770 Endopeptidase K Proteins 0.000 description 1
- 208000004232 Enteritis Diseases 0.000 description 1
- 241000588921 Enterobacteriaceae Species 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 1
- 102000016943 Muramidase Human genes 0.000 description 1
- 108010014251 Muramidase Proteins 0.000 description 1
- 108010062010 N-Acetylmuramoyl-L-alanine Amidase Proteins 0.000 description 1
- 108020002230 Pancreatic Ribonuclease Proteins 0.000 description 1
- 102000005891 Pancreatic ribonuclease Human genes 0.000 description 1
- 206010035664 Pneumonia Diseases 0.000 description 1
- 229920000294 Resistant starch Polymers 0.000 description 1
- 208000025865 Ulcer Diseases 0.000 description 1
- 241001394203 [Clostridium] leptum DSM 753 Species 0.000 description 1
- DPXJVFZANSGRMM-UHFFFAOYSA-N acetic acid;2,3,4,5,6-pentahydroxyhexanal;sodium Chemical compound [Na].CC(O)=O.OCC(O)C(O)C(O)C(O)C=O DPXJVFZANSGRMM-UHFFFAOYSA-N 0.000 description 1
- 230000000172 allergic effect Effects 0.000 description 1
- 230000009285 allergic inflammation Effects 0.000 description 1
- 230000000845 anti-microbial effect Effects 0.000 description 1
- 208000010668 atopic eczema Diseases 0.000 description 1
- 230000001363 autoimmune Effects 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 235000010948 carboxy methyl cellulose Nutrition 0.000 description 1
- 229920003123 carboxymethyl cellulose sodium Polymers 0.000 description 1
- 229940063834 carboxymethylcellulose sodium Drugs 0.000 description 1
- 230000024245 cell differentiation Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 239000000460 chlorine Substances 0.000 description 1
- 229910052801 chlorine Inorganic materials 0.000 description 1
- 230000004087 circulation Effects 0.000 description 1
- 210000001072 colon Anatomy 0.000 description 1
- 230000000112 colonic effect Effects 0.000 description 1
- 208000029742 colonic neoplasm Diseases 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 239000012154 double-distilled water Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 210000001842 enterocyte Anatomy 0.000 description 1
- 208000037902 enteropathy Diseases 0.000 description 1
- 210000000981 epithelium Anatomy 0.000 description 1
- 235000019441 ethanol Nutrition 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 235000013355 food flavoring agent Nutrition 0.000 description 1
- 239000003205 fragrance Substances 0.000 description 1
- 150000002431 hydrogen Chemical class 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 208000028774 intestinal disease Diseases 0.000 description 1
- 229960000274 lysozyme Drugs 0.000 description 1
- 239000004325 lysozyme Substances 0.000 description 1
- 235000010335 lysozyme Nutrition 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 235000012054 meals Nutrition 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 235000021140 nondigestible carbohydrates Nutrition 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 230000033116 oxidation-reduction process Effects 0.000 description 1
- 238000005554 pickling Methods 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 108700022487 rRNA Genes Proteins 0.000 description 1
- 235000021254 resistant starch Nutrition 0.000 description 1
- 238000005096 rolling process Methods 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 238000002864 sequence alignment Methods 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- MFBOGIVSZKQAPD-UHFFFAOYSA-M sodium butyrate Chemical compound [Na+].CCCC([O-])=O MFBOGIVSZKQAPD-UHFFFAOYSA-M 0.000 description 1
- 235000000346 sugar Nutrition 0.000 description 1
- 230000009897 systematic effect Effects 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
Classifications
-
- 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
- C12N1/205—Bacterial isolates
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/145—Clostridium
-
- 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
- A61K35/742—Spore-forming bacteria, e.g. Bacillus coagulans, Bacillus subtilis, clostridium or Lactobacillus sporogenes
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P5/00—Preparation of hydrocarbons or halogenated hydrocarbons
- C12P5/02—Preparation of hydrocarbons or halogenated hydrocarbons acyclic
- C12P5/023—Methane
-
- 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
- A61K2035/11—Medicinal preparations comprising living procariotic cells
- A61K2035/115—Probiotics
-
- 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
- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02E50/00—Technologies for the production of fuel of non-fossil origin
- Y02E50/30—Fuel from waste, e.g. synthetic alcohol or diesel
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Microbiology (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- General Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Medicinal Chemistry (AREA)
- Virology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biomedical Technology (AREA)
- Mycology (AREA)
- Molecular Biology (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Epidemiology (AREA)
- Pharmacology & Pharmacy (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a clostridium tender as well as a culture method and application thereof. The clostridium tender is extracted from the excrement of healthy people, and is classified and named as clostridium tender (c.), (clostridium leptum) L319 with deposit number GDMCC No: 60042. the invention also provides the clostridium tender (clostridium)clostridium leptum) Application of the culture method of L319. The culture method provided by the invention can improve the culture efficiency of clostridium tender. Clostridium tender (C.tenella) (C.tenella)clostridium leptum) The L319 has high enzyme activity, can degrade dietary fibers to generate butyrate, can induce the generation of immune suppressor cell Treg, and can also be used for generating hydrogen.
Description
Technical field
The present invention relates to one of biological field Clostridium leptum and its cultural method and applications.
Background technique
Clostridium leptum monoid is one of the three categories group that quantity is most dominant in human faecal mass flora, and Clostridium leptum is used as and is somebody's turn to do
The dominant bacteria of monoid, shared ratio is about 16% ± 7% in enteric bacteria, can be isolated from the excrement of Healthy People.
As probiotics, it plays an important role to our human health: first, promote the growth and differentiation of epithelial cell.
The butyric acid that Clostridium leptum generates can finally be metabolized and generate the proliferation of sodium butyrate inhibition colon epithelial cell system, promote colonic epithelium
The differentiation of cell line.Second, promote digestion and the Nutrition and Metabolism of food.Clostridium leptum has enzyme not available for human body (such as fibre
Tie up plain enzyme), it can be metabolized the unavailable Non-digestible carbohydrates of host itself, generate monosaccharide or short chain fatty acids by people
Body is utilized.Third adjusts the immune function of human body.Clostridium leptum can induce immunosuppressant cell Treg, can improve inflammation
Disease property enteropathy, including Crohn disease and ulcerative enteritis, while efficient treatment Allergic pulmonary inflammation, airway inflammation etc., greatly
The big immune function for maintaining human body.4th, the diversity of bacterium is adjusted, the stable state of enteric bacteria is maintained.5th, can ferment production
Raw acetic acid and hydrogen, acetic acid can be used as acidity regulator, acidulant, pickling agent, flavoring agent, fragrance etc., while be also good
Antimicrobial, and hydrogen has important purposes in industry and medical domain.Therefore, for the health care of human body, Clostridium leptum
With huge research significance.
Summary of the invention
It is an object of the present invention to provide a kind of Clostridium leptum, the clostridium dietary fiber that can degrade generates the prebiotic factor
Butyrate, clostridium itself can induce the generation of immunosuppressant cell Treg.
Clostridium provided by the present invention, be accredited as Clostridium leptum (Clostridium leptum), derive from Healthy People
The excrement of body.
The Clostridium leptum, classification naming be Clostridium leptum (Clostridium leptum) L319, it has been preserved in wide
East saves Culture Collection (abbreviation GDMCC, address: 5 building, the building of compound the 59th of Xianlie Middle Road, Guangzhou City 100), preservation
Number are as follows: GDMCC No:60042, preservation date are as follows: 25 months Mays in 2016.
Clostridium leptum provided by the present invention (Clostridium leptum) L319 has the feature that
1) colony characteristics: bacterium colony is rounded, and diameter is 0.2~0.5mm, white, surface bulge, the smooth of the edge.
2) cell morphological characteristic: cell is in the shape of a rod, slightly bent, and nearly terminal spore is seldom, both ends round blunt, and cell is about (1.3
~2.8) × (0.6~0.8) μm；Gram's staining is positive；.
3) physiological and biochemical property: obligate anaerobic growth；Not gelatin hydrolysate, indigestion haemocyanin being capable of metabolic resistance shallow lake
The polysaccharide such as powder, cellulose, hemicellulose, colloid and some oligosaccharides, resulting monosaccharide or short chain fatty acids can be by places
It is main to be utilized.
4) Clostridium leptum (Clostridium leptum) L319 16S rRNA gene order length be 1278bp, core
Nucleotide sequence is as shown in SEQ ID NO:1.
It is a further object to provide a kind of Clostridium leptum (Clostridium leptum) L319 culture side
Method.
Clostridium leptum provided by the present invention (Clostridium leptum) L319 cultural method, be by Clostridium leptum
(Clostridium leptum) L319 is inoculated in RCM culture medium, 32 DEG C ~ 38 DEG C, Anaerobic culturel under the conditions of pH 6 ~ 8.5.
Contain in 1000 mL RCM fluid nutrient mediums: tryptone, 10 g；Beef extract, 10 g；Glucose, 5 g；Chlorine
Change sodium, 5 g；Yeast extract, 3 g；Sodium acetate, 3 g；Soluble starch, 1 g；L-cysteine hydrochloride, 0.5 g；Surplus is to steam
Distilled water；
RCM solid medium is that addition 16g agar, distilled water are settled to 1000 mL into above-mentioned RCM culture medium.
Above-mentioned RCM culture medium further includes oxidation-reduction indicator resazurin, and content is 0.5 g/L.
In cultural method of the present invention, required anaerobic environment is by 80% N2、10% H2And 10%CO2Gaseous mixture composition,
General injection reaches the amount of 1 atmospheric pressure.
Culture Clostridium leptum provided by the present invention (Clostridium leptum) L319 method, culture efficiency is high,
Up to 4.67 × 109 CFU/mL。
Of the inventionIt is tenderClostridium (Clostridium leptum) L319 is isolated from the excrement of healthy human body
, it is the probiotics of human body intestinal canal, has overwhelming superiority on enterobacteriaceae monoid, account for about 16% ± 7%.The clostridium can degrade meals
The prebiotic factor butyrate that fiber generates needed by human body is eaten, butyrate is for the energy supply of human body and the development of enterocyte
It plays an important role, and inhibits the generation of colon cancer to a certain extent.In addition, the clostridium can also induce immunosuppressant cell
Treg, it is effective to treat inflammation and human autoimmune's property disease.Currently, just have research by way of oral Clostridium leptum come
Allergic inflammation, the airway inflammation for treating mouse.From this, Clostridium leptum (Clostridium leptum) L319 have it is wide
Wealthy research significance and background.
Strain of the present invention, classification naming be Clostridium leptum (Clostridium leptum) L319, preservation
In Guangdong Province's Culture Collection (GDMCC), deposit number are as follows: GDMCC No:60042, preservation date are as follows: 2016
May 25, preservation address are as follows: 5 building, the building of compound the 59th of Xianlie Middle Road, Guangzhou City 100.
Specific embodiment
Experimental method used in following embodiments is conventional method unless otherwise specified.
Basic composition is of RCM fluid nutrient medium used in experiment (/ liter):
Tryptone, 10 g；Beef extract, 10 g；Glucose, 5 g；Sodium chloride, 5 g；Yeast extract, 3 g；Sodium acetate, 3
g；Soluble starch, 1 g；L-cysteine hydrochloride, 0.5 g；Resazurin, 0.5 mg；Distilled water is settled to 1000 mL.
RCM solid medium used in experiment is the addition 16g agar into above-mentioned RCM fluid nutrient medium, makes its end
Concentration is 1.6%.
Embodiment 1, bacterial strain Clostridium leptum (Clostridium leptum) L319 separation preparation
The hydrolyzate for taking healthy human body excrement is prepared into outstanding bacterium solution, after be inoculated in RCM using resistant starch as substrate
Secondary culture is carried out in fluid nutrient medium, obtains enriched substance；Using enriched substance as inoculation source, it is inoculated in using cellulose the bottom of as
The RCM solid medium of object, is separated with Hungate rolling tube technique, finally obtains purifying strain.
It is preserved in Guangdong Province's Culture Collection (GDMCC), deposit number are as follows: GDMCC No:60042,
Preservation date are as follows: on May 25th, 2016, preservation address are as follows: 5 building, the building of compound the 59th of Xianlie Middle Road, Guangzhou City 100.
Embodiment 2, strain idenfication
One, form and physiological and biochemical property
By Clostridium leptum (Clostridium leptum) L319 is inoculated in the RCM solid culture using cellulose as substrate
Base is 7 in pH, and temperature is 37 DEG C, and gaseous mixture is 80% N2、10% H2And 10%CO2Under conditions of carry out 72 h of Anaerobic culturel.
The morphological feature of bacterium colony and cell is observed, and carries out following bio-chemical characteristics: 1 Gram stain test；2 liquid
Change gelatin test；The test of 3 haemocyanins；4 substrates utilize test.
It observes result and experimental result is as follows:
1) colony characteristics: bacterium colony is rounded, and diameter is 0.2~0.5mm, white, surface bulge, the smooth of the edge.
2) cell morphological characteristic: cell is in the shape of a rod, slightly bent, and nearly terminal spore is seldom, both ends round blunt, and cell is about (1.3
~2.8) × (0.6~0.8) μm；Gram's staining is positive.
3) physiological and biochemical property: obligate anaerobic growth；Not gelatin hydrolysate, indigestion haemocyanin being capable of metabolic resistance shallow lake
The polysaccharide such as powder, cellulose, hemicellulose, colloid and some oligosaccharides, resulting monosaccharide or short chain fatty acids can be by places
It is main to be utilized.
Two, Clostridium leptum (Clostridium leptum) L319 16S rRNA gene PCR amplification and sequencing:
1) extract DNA(and mention genomic kit referring to TAKARA):
By Clostridium leptum (Clostridium leptum) L319 is inoculated in RCM fluid nutrient medium and cultivated；Pass through survey
OD value takes the fermentation liquid for growing to late log phase, and 12,000 revs/min are centrifuged 5 minutes, removes supernatant；It is added 500 μ L's
Cell is resuspended in Buffer BS, and the Lysozyme(20 mg/mL of 50 μ L is added), it sufficiently inhales and plays mixing, incubate 1 in 37 DEG C of water-baths
Hour (was mixed by inversion primary) every 20 minutes；12,000 rpm are centrifuged 5 minutes, abandon supernatant；The Buffer of 180 μ L is added
The Proteinase K(20 mg/mL of GL, 20 μ L) and 10 μ L RNase A(10 mg/mL), sufficiently inhale play mixing, in 56
DEG C water-bath incubates 10 minutes；100% ethyl alcohol of the Buffer GB and 200 μ L of 200 μ L is added, sufficiently inhales and plays mixing；By Spin
Column is mounted on Collection Tube, and solution moves in Spin Column, and 12,000 rpm are centrifuged 2 minutes, abandons filter
Liquid；The Buffer WA of 500 μ L is added into Spin Column, 12,000 rpm are centrifuged 1 minute, abandon filtrate；By 700 μ L
Buffer WB be added into Spin Column, 12,000 rpm be centrifuged 1 minute, abandon filtrate, be repeated once；By Spin
Column is placed on Collection Tube, and 12,000 rpm are centrifuged 2 minutes；Spin Column is placed in new 1.5
On mL centrifuge tube, 65 DEG C of sterile purified waters of 70 μ L are added in the centre of Spin Column film, are stored at room temperature 5 minutes；12,
000 rpm is centrifuged 2 minutes eluted dnas.
2) PCR amplification and sequencing of 16S rRNA gene
Forward primer for PCR amplification is 5 '-CTGGCTTCGGGTATTACCGGCTCCC-3 ', reverse primer 5 '-
TTCAGAGGGGGATAACGTTCTGAAA-3'.PCR reaction system (25 μ L) are as follows: 12.5 μ L of Mix；1 μ L of forward primer；Instead
To 1 μ L of primer；2 μ L of DNA profiling；ddH2O 8.5 μL.PCR reaction condition are as follows: 95 DEG C of 5 min, 95 DEG C of 1 min, 60
DEG C 30 s, 72 DEG C of 1 min30 s, 72 DEG C of 7 min, 30 circulations, 4 DEG C of preservations.
The sequencing of PCR product is sent to Jin Weizhi company and is completed.
Sequencing result show Clostridium leptum (Clostridium leptum) the 16S rRNA gene order length of L319 is
1278bp, nucleotide sequence is as shown in SEQ ID NO:1.
16S rRNA gene order is subjected to sequence alignment analysis in GenBank, the results showed that, sequence withClostridium leptum16S rRNA gene order similitude be 100%.
To sum up, according to " Bergey ' s Manual of Systematic Bacteriology " (second edition), according to it
Morphological feature and physiological and biochemical property, and according to its comparison result of 16S rRNA gene order in GenBank, tender shuttle
Bacterium (Clostridium leptum) L319 be accredited as novel species Clostridium leptum (Clostridium leptum).
Embodiment 3, Clostridium leptum (Clostridium leptum) L319 culture
1) by Clostridium leptum (Clostridium leptum) L319 with 6% ratio is inoculated in 50 mL RCM Liquid Cultures
In base, cultivated under conditions of 27 DEG C, pH 6.0.
The composition (/ liter) of RCM fluid nutrient medium is as follows:
Tryptone, 10 g；Beef extract, 10 g；Glucose, 5 g；Sodium chloride, 5 g；Yeast extract, 3 g；Sodium acetate, 3
g；Soluble starch, 1 g；L-cysteine hydrochloride, 0.5 g；Resazurin, 0.5 mg；Distilled water is settled to 1000 mL.
Anaerobic environment needed for culture is by 80% N2、10% H2And 10%CO2Gaseous mixture composition, general injection reaches 1
The amount of a atmospheric pressure.
Experiment is repeated three times, and the mean concentration of thallus reaches (2.1 ± 0.4) × 109 CFU/mL。
2) by Clostridium leptum (Clostridium leptum) L319 with 6% ratio is inoculated in 50 mL RCM Liquid Cultures
Base is cultivated under conditions of 32 DEG C, pH 6.5.
The composition (/ liter) of RCM fluid nutrient medium is as follows:
Tryptone, 10 g；Beef extract, 10 g；Glucose, 5 g；Sodium chloride, 5 g；Yeast extract, 3 g；Sodium acetate, 3
g；Soluble starch, 1 g；L-cysteine hydrochloride, 0.5 g；Resazurin, 0.5 mg；Distilled water is settled to 1000 mL.
Required anaerobic environment is by 80% N2、10% H2And 10%CO2Gaseous mixture composition, general injection reaches 1 big
The amount of air pressure.
Experiment is repeated three times, and the mean concentration of thallus reaches (3.0 ± 0.4) × 109 CFU/mL。
3) by Clostridium leptum (Clostridium leptum) L319 with 6% ratio is inoculated in 50 mL RCM Liquid Cultures
Base is cultivated under conditions of 37 DEG C, pH 7.0.
The composition (/ liter) of RCM fluid nutrient medium is as follows:
Tryptone, 10 g；Beef extract, 10 g；Glucose, 5 g；Sodium chloride, 5 g；Yeast extract, 3 g；Sodium acetate, 3
g；Soluble starch, 1 g；L-cysteine hydrochloride, 0.5 g；Resazurin, 0.5 mg；Distilled water is settled to 1000 mL.
Required anaerobic environment is by 80% N2、10% H2And 10%CO2Gaseous mixture composition, general injection reaches 1 big
The amount of air pressure.
Experiment is repeated three times, and the mean concentration of thallus reaches (4.2 ± 0.4) × 109 CFU/mL。
4) by Clostridium leptum (Clostridium leptum) L319 with 6% ratio is inoculated in 50 mL RCM Liquid Cultures
Base is cultivated under conditions of 42 DEG C, pH 7.5.
The composition (/ liter) of RCM fluid nutrient medium is as follows:
Tryptone, 10 g；Beef extract, 10 g；Glucose, 5 g；Sodium chloride, 5 g；Yeast extract, 3 g；Sodium acetate, 3
g；Soluble starch, 1 g；L-cysteine hydrochloride, 0.5 g；Resazurin, 0.5 mg；Distilled water is settled to 1,000 mL.
Required anaerobic environment is by 80% N2、10% H2And 10%CO2Gaseous mixture composition, general injection reaches 1 big
The amount of air pressure.
Experiment is repeated three times, and the mean concentration of thallus reaches (3.3 ± 0.4) × 109 CFU/mL。
5) by Clostridium leptum (Clostridium leptum) L319 with 6% ratio is inoculated in 50 mL RCM Liquid Cultures
Base is cultivated under conditions of 47 DEG C, pH 8.0.
The composition (/ liter) of RCM fluid nutrient medium is as follows:
Tryptone, 10 g；Beef extract, 10 g；Glucose, 5 g；Sodium chloride, 5 g；Yeast extract, 3 g；Sodium acetate, 3
g；Soluble starch, 1 g；L-cysteine hydrochloride, 0.5 g；Resazurin, 0.5 mg；Distilled water is settled to 1000 mL.
Required anaerobic environment is by 80% N2、10% H2And 10%CO2Gaseous mixture composition, general injection reaches 1 big
The amount of air pressure.
Experiment is repeated three times, and the mean concentration of thallus reaches (2.3 ± 0.3) × 109 CFU/mL。
6) by Clostridium leptum (Clostridium leptum) L319 with 6% ratio is inoculated in 50 mL RCM Liquid Cultures
Base is cultivated under conditions of 47 DEG C, pH 8.5.
The composition (/ liter) of RCM fluid nutrient medium is as follows:
Tryptone, 10 g；Beef extract, 10 g；Glucose, 5 g；Sodium chloride, 5 g；Yeast extract, 3 g；Sodium acetate, 3
g；Soluble starch, 1 g；L-cysteine hydrochloride, 0.5 g；Resazurin, 0.5 mg；Distilled water is settled to 1000 mL.
Required anaerobic environment is by 80% N2、10% H2And 10%CO2Gaseous mixture composition, general injection reaches 1 big
The amount of air pressure.
Experiment is repeated three times, and in repeating three times, thallus is not grown.
The results showed that Clostridium leptum (Clostridium leptum) L319 progress anaerobic growth, at 27 DEG C -42 DEG C
At a temperature of strain growth, do not grown at 47 DEG C, the optimum growth temperature of bacterial strain is 32 DEG C -38 DEG C；PH growth scope 6-8.5,
The most suitable growth pH is 6.5-7.0.Under optimum growing condition, cell concentration is up to (4.2 ± 0.4) × 109CFU/mL。
Embodiment 4, Clostridium leptum (Clostridium leptum) L319 cellulase enzyme activity determination
One, enzyme solution is prepared
By Clostridium leptum (Clostridium leptum) L319 is inoculated in RCM fluid nutrient medium, pH 7, cultivation temperature
It is 37 DEG C, after cultivating 72h, centrifugation removes supernatant；Cell precipitation is resuspended with PBS buffer solution, obtains enzyme solution.
RCM culture medium group becomes (/ liter):
Tryptone, 10 g；Beef extract, 10 g；Glucose, 5 g；Sodium chloride, 5 g；Yeast extract, 3 g；Sodium acetate, 3
g；Soluble starch, 1 g；L-cysteine hydrochloride, 0.5 g；Resazurin, 0.5 mg；Distilled water is settled to 1000 mL.
Required anaerobic environment is by 80% N2、10% H2And 10%CO2Gaseous mixture composition, general injection reaches 1 big
The amount of air pressure.
Two, the drafting of standard curve
1) 0.2,0.4,0.6,0.8,1.0mL glucose is drawn respectively in colorimetric cylinder, is diluted to distilled water
1mL adds 3.5- dinitrosalicylic acid color developing agent 3mL, and colour developing 10 minutes is boiled in boiling water bath, cooling, adds distilled water 21mL, shakes
It is even.
2) sugar is replaced to make blank tube with 1mL distilled water, the colorimetric at 550nm.It is micro- with glucose using optical density as ordinate
G number is abscissa, draws standard curve.
Three, the measurement of blank tube
1mL enzyme solution is taken, boiling water bath 5 minutes, cooling plus 3Ml 0.5%CMC was put into 50 DEG C of water-baths 30 simultaneously with sample cell and divides
Clock.Other same sample cells of operation.
Four, the measurement of sample
0.5% carboxymethylcellulose sodium solution 3mL, enzyme solution 1mL are taken, is saccharified 30 minutes in 50 DEG C of water-baths, is taken out, immediately
Boiling in boiling water bath 10 minutes inactivates enzyme, obtains saccharified liquid, cooling that 3mL developing solution, then boiling water bath 10 minutes are added, after cooling
Add water to be settled to 25mL, mix, 550nm surveys OD value.
Three repeated experiments are carried out respectively, are averaged.
Compared to CM slant medium and PY- maltose broth bouillon culture Clostridium leptum ATCC 29065 is used, make
With the Clostridium leptum of RCM culture medium culture (Clostridium leptum) L319 enzyme activity is higher, it is 2450 U/mL.
Embodiment 5, Clostridium leptum (Clostridium leptum) measurement of hydrogen in L319
Hydrogen abundant can be generated in Clostridium leptum, and hydrogen has important purposes in industry and medical domain.
By Clostridium leptum (Clostridium leptum) L319 is inoculated in 50 mL RCM fluid nutrient mediums, at 37 DEG C,
It is cultivated under conditions of pH 7.0.
RCM culture medium group becomes (/ liter):
Tryptone, 10 g；Beef extract, 10 g；Glucose, 5 g；Sodium chloride, 5 g；Yeast extract, 3 g；Sodium acetate, 3
g；Soluble starch, 1 g；L-cysteine hydrochloride, 0.5 g；Resazurin, 0.5 mg；Distilled water is settled to 1000 mL.
Required anaerobic environment is by 80% N2、10% H2And 10%CO2Mixed gas composition, general injection reaches 1
The amount of atmospheric pressure.
Experimental provision selects the sampler bag of E-Switch and the gas-phase apparatus of Thermo.
One, the acquisition of gas
Acquire the gas of 10 bottles of fluid nutrient mediums, sealing simultaneously using collection bag.
Two, the detection of hydrogen
1) H by rotating sampling valve V1, in sample2, O2, N2, CO2Component enters Porapak N chromatographic column quickly.
2) in Porapak N chromatographic column, CO2It is kept completely separate with other components, other non-separation components enter
MolSieve 5A molecular sieve chromatography column.
3) H2It first flows out from MolSieve 5A molecular sieve chromatography column, is detected into TCD detector.
4) work as H2After from the outflow of MolSieve 5A molecular sieve chromatography column into detector, rotation V2 to ON state, quilt
Isolated CO2Component is directly entered detector detection without MolSieve 5A chromatographic column.
5) work as CO2Into after detector, V2, V1 switch to the state of OFF, CO2Subsequent component is gone out by blowback, by every
From the O in molecular sieve chromatography column2, N2, component obtains being segregated into detector detection.
6) V1 and V2 returns back to original state OFF, and preparing next time, sample introduction is analyzed.
By duplicate experiment three times, finally analyze hydrogen yield be 3.84 × 105~4.61×105ppm。
Sequence table
<110>Nanjing University of Technology
<120>a kind of Clostridium leptum and its cultural method and application
<130> xb16082203
<160> 1
<170> PatentIn version 3.5
<210> 1
<211> 1278
<212> DNA
<213> Clostridium leptum
<400> 1
ctggcttcgg gtattaccgg ctcccatggt gtgacgggcg gtgtgtacaa ggcccgggaa 60
cgtattcacc gcggcatgct gatccgcgat tactagcaat tccgacttca cgcaggcggg 120
ttgcagcctg cgatccgaac tgggactgct tttagggttt tgctccacct cgcggctttg 180
cttccctctg tttacagcca ttgtagtacg tgtgtggccc aggtcataag gggcatgatg 240
atttgacgtc gtccccacct tcctccgttt tgtcaacggc agtctctcta gagtgctctt 300
gcgtagcaac tagaaacaag ggttgcgctc gttgcgggac ttaacccaac atctcacgac 360
acgagctgac gacaaccatg caccacctgt ctcgcctttc cccgaagggc acctaatgca 420
tctctgcttc gttagacgga tgtcaagacc tggtaaggtt cttcgcgttg cttcgaatta 480
aaccacatac tccactgctt gtgcgggccc ccgtcaattc ctttgagttt caaccttgcg 540
gccgtactcc ccaggtggat tacttattgt gttaactgcg gcacggaggg ggtcagaccc 600
cccacaccta gtaatcatcg tttacggcat ggactaccag ggtatctaat cctgtttgct 660
acccatgctt tcgtgcttca gcgtcagtta aagcccagta ggccgccttc gccactggtg 720
ttcctcccga tctctacgca tttcaccgct acaccgggaa ttccgcctac ctctacttca 780
ctcaagcaag acagtttcaa gcgcagttta tgggttaagc ccatagattt cacgcctgac 840
ttgcctcgcc gcctacgcac cctttacacc cagtaaatcc ggacaacgct cgctccctac 900
gtattaccgc ggctgctggc acgtagttag ccggagcttt ctccttagct accgtcatta 960
tcgtcactaa gaacagaggt ttacaatccg aaaaccttct tccctcacgc ggcgttgctg 1020
catcagggtt tcccccattg tgcaatatcc cccactgctg cctcccgtag gagtctgggc 1080
cgtgtctcag tcccaatgtg gccgttcaac ctctcagtcc ggctaccgat cgtcgctttg 1140
gtgggccgtt accccgccaa ctggctaatc ggacgcgagt ccatcttcca gcggattgct 1200
cctttgatca acctatcatg cgataaattg atgttatgcg gtattagcgt tcttttcaga 1260
acgttatccc cctctgaa 1278
Claims (7)
1. a kind of Clostridium leptum, which is characterized in that its classification naming be Clostridium leptum (Clostridium leptum) L319,
Deposit number is GDMCC No:60042.
2. the cultural method of Clostridium leptum described in claim 1, which is characterized in that by Clostridium leptum (Clostridium leptum) L319 is inoculated in RCM culture medium, 32 DEG C ~ 38 DEG C, Anaerobic culturel under the conditions of pH 6 ~ 8.5；
The RCM culture medium is using cellulose as substrate；
RCM Liquid Culture based component are as follows: tryptone, 10 g；Beef extract, 10 g；Glucose, 5 g；Sodium chloride, 5 g；Yeast
Cream, 3 g；Sodium acetate, 3 g；Soluble starch, 1 g；L-cysteine hydrochloride, 0.5 g；Distilled water is settled to 1000 mL；
RCM solid medium is that addition 16g agar, distilled water are settled to 1000 mL into above-mentioned RCM fluid nutrient medium.
3. according to the method described in claim 2, it is characterized in that, containing 0.5 g/L resazurin in the RCM culture medium.
4. according to the method described in claim 2, it is characterized in that, the anaerobic environment is made of anaerobic gas, the gas
Body group is divided into 80% N2、10% H2And 10%CO2。
5. according to the method described in claim 2, it is characterized in that, the condition of culture be 37 DEG C, pH 7, Anaerobic culturel
72h。
6. application of the Clostridium leptum described in claim 1 in hydrogen producing.
7. application of the Clostridium leptum described in claim 1 in enteron aisle adjusting decomposes dietary fiber and generate the prebiotic factor, treatment is scorching
Disease and autoimmune disease adjust intestinal microecology balance.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610698863.XA CN106167778B (en) | 2016-08-22 | 2016-08-22 | Clostridium tender and culture method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610698863.XA CN106167778B (en) | 2016-08-22 | 2016-08-22 | Clostridium tender and culture method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106167778A CN106167778A (en) | 2016-11-30 |
| CN106167778B true CN106167778B (en) | 2019-09-03 |
Family
ID=57376622
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610698863.XA Active CN106167778B (en) | 2016-08-22 | 2016-08-22 | Clostridium tender and culture method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106167778B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108034621A (en) * | 2018-01-31 | 2018-05-15 | 安徽瑞思威尔科技有限公司 | A kind of enrichment method of pit mud anaerobism clostridium Clostridium |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2012142605A1 (en) * | 2011-04-15 | 2012-10-18 | Samaritan Health Services | Rapid recolonization deployment agent |
| CN104357496B (en) * | 2014-10-29 | 2017-06-27 | 中国科学院成都生物研究所 | A kind of method by microorganism catalysis lactic acid synthesizing hexanoic acid |
| CN104630292A (en) * | 2015-02-09 | 2015-05-20 | 哈尔滨工业大学宜兴环保研究院 | Method for preparing butyric acid by fermenting lignocellulose by using mixed flora |
-
2016
- 2016-08-22 CN CN201610698863.XA patent/CN106167778B/en active Active
Non-Patent Citations (1)
| Title |
|---|
| Development and application of a functional CE-SSCP fingerprinting method based on [Fe-Fe]-hydrogenase genes for monitoring hydrogen-producing Clostridium in mixed cultures;Marianne，et al;《International Journal of Hydrogen Energy》;20101231;第35卷(第24期);全文 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106167778A (en) | 2016-11-30 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Macfarlane et al. | Colonization of mucin by human intestinal bacteria and establishment of biofilm communities in a two-stage continuous culture system | |
| US10113145B2 (en) | Paenibacillus sp. strain, cultivation method and use of the same | |
| Canale-Parola | Biology of the sugar-fermenting Sarcinae | |
| Nicolaus et al. | Haloarcula spp able to biosynthesize exo-and endopolymers | |
| CN102533588B (en) | Lactobacillus brevis for producing extracellular exopolysaccharide and application thereof | |
| Miranda-Tello et al. | Garciella nitratireducens gen. nov., sp. nov., an anaerobic, thermophilic, nitrate-and thiosulfate-reducing bacterium isolated from an oilfield separator in the Gulf of Mexico | |
| Montgomery et al. | Characterization of rat cecum cellulolytic bacteria | |
| CN105969681A (en) | Highly oxidation-resistant, cholate-tolerant and acid-resistant lactobacillus plantarum JR4 and application thereof | |
| CN102703339A (en) | High-yield arginine deiminase bacterial strain and method for producing L-citrulline by same | |
| CN104046586B (en) | One strain gene engineering bacterium and the application in producing (2R, 3R)-2,3-butanediol thereof | |
| CN106167778B (en) | Clostridium tender and culture method and application thereof | |
| CN104877942B (en) | Series bacillus novel bacterial and its cultural method and application | |
| CN112795492A (en) | Isaria cicadae strain capable of producing exopolysaccharide, preparation method and application | |
| CN108220189A (en) | One plant of marine bacteria for producing zytase and application | |
| CN105586293B (en) | A kind of new lactic acid utilizes clostridium and application thereof | |
| CN103114053B (en) | Produce Lactobacillus coryniformis Shaanxi subspecies and the application thereof of high optical purity D-lactic acid | |
| WO2021056683A1 (en) | Strain for producing lipase and application thereof | |
| CN103468606B (en) | Klebsiella oxytoca and application thereof in allitol production | |
| CN115927023A (en) | Abnormal yeast Weikehan and application thereof, and wolfberry wine and preparation method thereof | |
| CN106479925B (en) | A kind of Klebsiella for producing beta galactosidase and its application | |
| CN109456898A (en) | A kind of the fermentation preparation and its application of chaetomium globosum dextranase | |
| CN112094762B (en) | Corynebacteria vinifera strain and application thereof | |
| TWI714258B (en) | Preparation method for improving phycoerythrin yield | |
| CN107267420B (en) | High-yield β -1, 3-glucan bacillus subtilis and application thereof | |
| Zeng et al. | Development of new strategies for the production of high-purity fructooligosaccharides using β-fructofuranosidase and a novel isolated Wickerhamomyces anomalus |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |