WO2017033925A1 - Bactérie productrice d'acide butyrique - Google Patents

Bactérie productrice d'acide butyrique Download PDF

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WO2017033925A1
WO2017033925A1 PCT/JP2016/074497 JP2016074497W WO2017033925A1 WO 2017033925 A1 WO2017033925 A1 WO 2017033925A1 JP 2016074497 W JP2016074497 W JP 2016074497W WO 2017033925 A1 WO2017033925 A1 WO 2017033925A1
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butyric acid
acid
producing bacterium
yit
strain
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PCT/JP2016/074497
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Japanese (ja)
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敏彦 高田
久代　明
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株式会社ヤクルト本社
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Priority to JP2017536440A priority Critical patent/JP6712598B2/ja
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K10/00—Animal feeding-stuffs
    • A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
    • A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
    • A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
    • 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
    • A61K35/742—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
    • 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
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/40—Preparation of oxygen-containing organic compounds containing a carboxyl group including Peroxycarboxylic acids
    • C12P7/52—Propionic acid; Butyric acids
    • 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

Definitions

  • the present invention relates to a new butyric acid-producing bacterium and a butyric acid production enhancer containing the same.
  • Short-chain fatty acids produced by intestinal bacteria lower the pH in the intestine, suppress the growth of harmful bacteria, and are rapidly absorbed from the intestinal epithelium to be used as a host energy source via the portal vein Yes.
  • butyric acid is the most important energy source of intestinal epithelial cells, and is considered to have beneficial effects on the host, such as promoting mucin secretion via the vagus nerve.
  • Blautia coccoides group and Clostridium leptum ⁇ ⁇ subgroup which are the most dominant bacteria in the human intestine, contain many bacterial species that produce butyric acid and play an important role in maintaining the intestinal environment of the host.
  • a decrease in Blautia coccoides group and Clostridium leptum subgroup and a concomitant decrease in the butyric acid concentration in the intestinal tract have been reported (Non-patent Document 1).
  • Faecalibacterium prausnitzii a major butyrate-producing bacterium belonging to Clostridium leptum subgroup, induces the expression of anti-inflammatory cytokine IL-10 in an experimental system using cultured cells, and is administered to inflammatory bowel disease model mice. Is also attracting attention as one of the next-generation probiotic candidates (Non-Patent Document 2).
  • Vermeiren J. et al. 2012, FEMS Microbiol. Ecol. Sokol, H. et al. 2008, Proc Natl Acad Sci USA.
  • bacteria belonging to Clostridium leptum subgroup are bacteria that cannot produce butyric acid in a medium without acetic acid or that also produce formic acid together with butyric acid. It is known that the amount of short-chain fatty acids such as acetic acid and butyric acid is lower in patients with inflammatory bowel disease than in healthy individuals. Is useful in view of the beneficial effects of butyric acid in the intestine.
  • formic acid is corrosive and is also known as a mitochondrial toxin that can inhibit cytochrome c oxidase (Patent Document 1), and it is not preferable to ingest bacteria that produce formic acid.
  • an object of the present invention is to provide a new butyric acid-producing bacterium that produces butyric acid even in the absence of acetic acid and does not produce formic acid, and a butyric acid production enhancer containing the bacterium.
  • the present inventor tried to isolate a new butyric acid-producing bacterium from the sample library owned by the applicant, and as a result, produced butyric acid even in the absence of acetic acid, and further produced Clostridium leptum subgroup, which does not produce formic acid.
  • a new butyric acid-producing bacterium belonging to the present invention was found and the present invention was completed.
  • the present invention provides the following [1] to [7].
  • [1] A butyric acid-producing bacterium belonging to Clostridium leptum subgroup, which produces butyric acid and does not produce formic acid in a medium without acetic acid.
  • the number of bacteria is 1.0 ⁇ 10 6 cells inoculated into 2 mL of acetic acid-free YCG medium, and the amount of butyric acid produced when anaerobically cultured at 37 ° C. for 72 hours is 10 mM or more, and the amount of formic acid produced is 1 mM.
  • a butyric acid production enhancer comprising the butyric acid-producing bacterium according to any one of [1] to [4].
  • a composition for food, beverage, medicine or feed containing the butyric acid-producing bacterium according to any one of [1] to [4].
  • the butyric acid-producing bacterium of the present invention is a new bacterial species, and produces butyric acid even in the absence of acetic acid, and does not produce formic acid. Therefore, it is highly safe and is used as a butyric acid production enhancer in the intestine. It is useful as a product, medicine, and feed composition.
  • the butyric acid-producing bacterium of the present invention is a bacterium that produces butyric acid and does not produce formic acid in a medium without acetic acid, and belongs to the C. leptum subgroup.
  • the butyric acid-producing bacterium belonging to the conventional C. leptum subgroup is a bacterium that cannot produce butyric acid in the absence of acetic acid or that produces formic acid together with butyric acid
  • the butyric acid-producing bacterium of the present invention is a new one. It is a new bacterial species with characteristics.
  • the absence of acetic acid means that acetic acid is not present, or even if it is present, only a trace amount of 1 mM or less is present.
  • not producing formic acid means that formic acid is not produced at all or even when produced, it is less than 1 mM.
  • the characteristics of the butyric acid-producing bacterium of the present invention are inoculated with 2 mL of YCG (yeast extract-casitone glucose) medium in the absence of acetic acid at 1.0 ⁇ 10 6 cells and anaerobically cultured at 37 ° C. for 72 hours.
  • the butyric acid production amount is preferably 10 mM or more, and the formic acid production amount is preferably less than 1 mM, the butyric acid production amount under the conditions is 10 to 30 mM, and the formic acid production amount is less than 0.7 mM. More preferably.
  • the strain frozen stock solution is preferably a solution in which the cells are suspended in a 10% by mass skim milk medium or 20% by mass glycerol-added Mueller Hinton medium.
  • the measurement of the concentration of organic acids such as butyric acid and formic acid in the culture solution is not particularly limited as long as the organic acid concentration can be measured. For example, it can be measured by an HPLC system for organic acid analysis.
  • the butyric acid-producing bacterium of the present invention produces butyric acid even in a medium containing acetic acid.
  • butyric acid-producing bacterium of the present invention examples include Butyricicoccus sp. YIT 12787 (NITE BP-02106), Butyricicoccus sp. YIT 12788 (NITE BP-02095) and their mutants.
  • YIT 12787 and YIT 12788 are registered in the Patent Microorganisms Depositary Center of the National Institute of Technology and Evaluation on August 20, 2015 and July 31, 2015. Deposited each day.
  • the 16S rRNA of YIT 12787 was SEQ ID NO: 3
  • the 16S rRNA of YIT 12788 was SEQ ID NO: 4.
  • Examples of the mutant strains of YIT 12787 and YIT 12788 in the present invention include mutants having a homology of 16S rRNA of 99% or more with SEQ ID NO: 3 and / or SEQ ID NO: 4. In the present invention, homology refers to the identity of base sequences.
  • the homology between YIT 12787 and YIT 12788 and related species was examined, the homology between YIT 12787 and Eubacterium desmolans ATCC 43058 T was 7.5 to 18.9%, and YIT 12787 and Butyricicoccus pullicaecorum 25 The homology with ⁇ 3 T was 10.3-14.3%. Further, the homology between YIT 12788 and Eubacterium desmolans ATCC 43058 T was 7.9 to 17.8%, and the homology between YIT 12788 and Butyricicoccus pullicaecorum 25-3 T was 10.7 to 16.5%. . Therefore, the butyric acid-producing bacterium of the present invention has a DNA homology of 20% or less with the related species Eubacterium desmolans ATCC 43058 T and Butyricicoccus pullicaecorum 25-3 T.
  • the phylogenetic tree of the butyric acid-producing bacterium of the present invention is as shown in FIG. 1, which is a new bacterium species belonging to C. leptum subgroup.
  • the butyric acid-producing bacteria of the present invention include bacteria having the following properties (1) and (2).
  • (1) The homology between the base sequence of 16S rRNA and SEQ ID NO: 3 and / or SEQ ID NO: 4 is 99% or more.
  • DNA homology with the related species Eubacterium desmolans ATCC 43058 T and Butyricicoccus pullicaecorum 25-3 T is 20% or less.
  • the butyric acid-producing bacterium of the present invention has the following mycological properties. As shown in Examples below, the butyric acid-producing bacterium of the present invention is different from Eubacterium desmolans ATCC 43058 T and Butyricicoccus pullicaecorum 25-3 T in sugar fermentation properties and enzyme activity properties. It can be seen that this is a new species belonging to the leptum subgroup.
  • the butyric acid-producing bacterium of the present invention is inoculated and cultured with various sample libraries and microorganisms collected from nature, and the grown colonies are fished and measured for the ability to produce butyric acid and formic acid in a medium without acetic acid. Can be isolated.
  • the confirmation of the butyric acid-producing bacterium of the present invention can be confirmed by organic acid analysis, sequence analysis, or the like.
  • the butyric acid-producing bacterium of the present invention can be proliferated and subcultured by anaerobic culture using a GAM medium or PY (peptone yeast extract) medium prepared in an anaerobic state, for example.
  • GAM medium or PY (peptone yeast extract) medium prepared in an anaerobic state, for example.
  • the butyric acid-producing bacterium of the present invention has no safety problem, produces butyric acid in the absence of acetic acid, and does not produce formic acid. Therefore, the composition containing the butyric acid-producing bacterium of the present invention is useful as a composition for foods and drinks, medicines or feeds.
  • the composition is useful as a butyric acid production enhancer because it produces butyric acid in the intestines of animals including humans.
  • butyric acid is not only used as an energy source for colonic mucosal epithelial cells, but also has an epithelial cell growth promoting action, an anti-inflammatory action, and an intestinal motility enhancing action, as well as colon cancer and ulcerative colitis. Therefore, the composition of the present invention is particularly useful as a pharmaceutical, food, drink or feed having these physiological activities.
  • composition of the present invention preferably contains 10 4 cfu to 10 14 cfu of butyric acid-producing bacteria as viable bacteria.
  • composition of the present invention can be in a form suitable for each of foods and drinks, medicines and feeds.
  • a medicine for example, it can be mixed with a solid or liquid non-toxic pharmaceutical carrier to form a conventional pharmaceutical preparation.
  • preparations include solid preparations such as tablets, granules, powders and capsules, liquid preparations such as solutions, suspensions and emulsions, and freeze-dried preparations. These preparations can be prepared by conventional means on the preparation.
  • non-toxic pharmaceutical carrier examples include glucose, lactose, sucrose, starch, mannitol, dextrin, fatty acid glyceride, polyethylene glycol, hydroxyethyl starch, ethylene glycol, polyoxyethylene sorbitan fatty acid ester, amino acid, gelatin, albumin , Water, physiological saline and the like.
  • conventional additives such as stabilizers, wetting agents, emulsifiers, binders, isotonic agents, excipients and the like can be appropriately added as necessary.
  • an additive that can be used as a food or drink is appropriately used, and it may be formed into a form suitable for food, that is, a granule, a granule, a tablet, a capsule, a paste, or the like, using a conventional means.
  • the types of food and drink include, for example, processed meat foods such as ham and sausage, processed fishery foods such as kamaboko and chikuwa, foods such as bread, confectionery, butter and powdered milk, water, fruit juice, milk, soft drinks and tea drinks. And the like. The same applies to feed.
  • composition of the present invention When the composition of the present invention is administered to animals including humans, it is preferably administered orally or enterally from the viewpoint of producing butyric acid in the intestine, and its dosage is as a butyric acid producing bacterium per day.
  • the number of viable bacteria is preferably 1.0 ⁇ 10 4 cfu or more, and more preferably 1.0 ⁇ 10 8 cfu to 1.0 ⁇ 10 12 cfu.
  • Example 1 Isolation of Butyrate-Producing Bacteria Two types (A, B) of samples were selected from the sample library owned by the applicant, and YCFA-M medium (YCFA supplemented with 0.5% mucin (acetic acid 20 mM, propionic acid) was selected. 1 mL of 5 mM) medium) was inoculated with 10 ⁇ L of a 10-fold diluted solution of each sample. After anaerobic culture at 37 ° C. for 8 hours, 10 ⁇ L of the supernatant after centrifugation at 4,000 G for 5 minutes was inoculated into 2 mL of new YCFA-M medium.
  • YCFA-M medium YCFA supplemented with 0.5% mucin (acetic acid 20 mM, propionic acid) was selected. 1 mL of 5 mM) medium
  • mucin acetic acid 20 mM, propionic acid
  • Example 2 Butyric acid and formic acid producing ability of strain A and strain B (1) Comparison of butyric acid and formic acid producing ability with known strains For strain A and strain B, including bacteria belonging to C. leptum subgroup in a medium without acetic acid The amount of butyric acid produced was compared with 18 known butyric acid producing bacteria (Table 1).
  • the supernatant after centrifugation was filtered through a 0.45 ⁇ m filter, and the amount of organic acid in the filtrate was measured using an organic acid analysis HPLC system described later.
  • organic acid analysis HPLC system There are 9 types of standard substances: succinic acid, sodium formate, sodium acetate, sodium propionate, sodium isobutyrate, n-sodium butyrate, sodium isovalerate, n-sodium valerate and lithium lactate (all Kanto Chemical Co., Ltd.)
  • Using a mixed aqueous solution of HPLC grade manufactured by the company quantification was performed by a two-point (0.01 and 0.2 ⁇ mol / 10 ⁇ L) absolute calibration curve method.
  • Table 2 shows the results of the organic acid producing ability of strain A and strain B using various media.
  • Strains A and B produced butyric acid in excess of 10 mM in the M2GSC medium without acetic acid, while the amount of formic acid produced was less than 1 mM and did not produce formic acid. Further, 20 mM or more of butyric acid was produced in the M2GSC + SCFA and YCFA medium supplemented with acetic acid. In the M2GSC + SCFA and YCFA media, the concentration of acetic acid after culturing was significantly lower than the initial content (Table 2), so it was considered that butyric acid was produced using acetic acid in the media. .
  • Example 3 16S rRNA sequence analysis of strain A and strain B
  • DNA was extracted by the bead phenol method. That is, the operation of suspending 200 ⁇ L of the bacterial solution of strain A and strain B in 1.0 mL of PBS, centrifuging at 15,000 rpm, and discarding the supernatant was repeated three times. The obtained pellet was first suspended in 450 ⁇ L of Extraction Buffer (100 mM Tris-HCl, 40 mM EDTA, pH 9.0) and 50 ⁇ L of 10% SDS, 0.3 g of glass beads (diameter 0.1 mm) and 500 ⁇ L of TE saturated.
  • Extraction Buffer 100 mM Tris-HCl, 40 mM EDTA, pH 9.0
  • Phenol was added and shaken vigorously for 30 seconds with FastPrep FP120 (power level 5.0). After centrifugation at 15,000 rpm for 10 minutes, isopropanol precipitation was performed, and the obtained DNA was dissolved in 1,000 ⁇ L of TE buffer. The obtained DNA solution was diluted 10 times with purified water to obtain a template DNA solution. Using the 8F and 15R primers shown in Table 3 for the template DNA solution, after reacting at 95 ° C for 5 minutes, 95 cycles of 95 ° C for 30 seconds, 55 ° C for 30 seconds, 72 ° C for 1 minute 30 seconds, 35 cycles The PCR reaction was performed, followed by a reaction at 72 ° C. for 5 minutes.
  • the PCR product was purified using High Pure PCR Product Purification Kit (Roche) and subjected to a sequencing reaction using BigDye Terminator v3.1 Cycle Sequence Kit (Applied Biosystems). The obtained sequence was subjected to a BLAST search of the Japan DNA Data Bank (DDBJ) and collated with a sequence database of known bacterial species. Furthermore, phylogenetic analysis was performed on the sequence of the isolates by the neighbor joining (NJ) method using Clustal W, and a phylogenetic tree was created using the Tree-View program.
  • DDBJ Japan DNA Data Bank
  • NJ neighbor joining
  • the created phylogenetic tree is shown in FIG.
  • the strain A was 1,495 bp (SEQ ID NO: 3) and the strain B was 1,500 bp (SEQ ID NO: 4). Both strains had 99% or more homology with each other.
  • strain A and strain B belonged to C. leptum subgroup and were considered to be closely related to Eubacterium desmolans ATCC 43058 T and Butyricicoccus pullicaecorum 25-3 T , which are known bacterial species.
  • strain A and Eubacterium desmolans ATCC 43058 T were 94.9%, and the homology between Butyricicoccus pullicaecorum 25-3 T was 96.3%.
  • the homology between strain B and Eubacterium desmolans ATCC 43058 T was 95.0%, and the homology between Butyricicoccus pullicaecorum 25-3 T was 96.3%.
  • the homology of 16S rRNA is 99% or more, but since strain A and strain B have less than 99% homology with known strains, new strains belonging to C. leptum subgroup It was thought that.
  • Strain A was transformed into Butyricicoccus sp.
  • YIT 12787 strain B was transformed into Butyricicoccus sp. It was named YIT 12788 and the properties of these strains belonging to the new strain were confirmed.
  • Example 4 Various biochemical properties
  • strains used were Eubacterium desmolans ATCC 43058 T (E. desmolans) and Butyricicoccus pullicaecorum 25-3 T (B. pulllicaecorum). The result was determined visually after 30 hours inoculation with the bacterial solution.
  • the results of the sugar fermentation property test are shown in Table 4.
  • YIT 12787 and YIT 12788 were positive for glucose utilization. All of E. desmolans were negative.
  • B.pullicaecorum assimilation of glucose, saccharose, salicin and xylose and hydrolysis of esculin were positive.
  • YIT 12787 and YIT 12788 can be distinguished from E. desmolans by the availability of glucose and the presence or absence of alkaline phosphatase, esterase (C4) and leucine allylamidase activity. Met. Moreover, it was distinguishable from B.pullicaecorum by the availability of saccharose, salicin and xylose, the hydrolytic ability of esculin, and the presence or absence of esterase (C4) activity.
  • the cells after fixation were washed 3 times with 5 mM EDTA solution, resuspended in 6 mL of the same solution, 1 mL each of 600 U / mL achromopeptidase and 50 mg / mL lysozyme were added, and the cells were lysed at 37 ° C. for 2 hours. .
  • 0.6 mL of SDS solution was added and treated at 60 ° C. for 10-30 minutes. Further, 150 ⁇ L of 10 mg / mL proteinase K was added and treated at 65 ° C.
  • a sequence library was prepared using TruSeq DNA PCR-Free Sample Prep LS Kit (Illumina) and DNA Shearing System M220 (Covaris) according to the protocol provided by Illumina.
  • the prepared sequence library was subjected to a 250 bp ⁇ 2 paired end sequence by the next-generation sequencer MiSeq (Illumina), the base sequence was decoded, and GC% was calculated based on the obtained base sequence result.
  • the GC content of the bacterial DNA was 53.9% for YIT 12787, 54.1% for YIT 12788, 54.3% for E. desmolans, and 54.0% for B.pullicaecorum. It was.
  • YIT 12787 and YIT 12788 have the following common biochemical properties.
  • Example 5 Properties of New Isolate One strain (C) was selected from the sample library owned by the applicant, and strain C was isolated by the method described in Example 1.
  • strain C and YIT 12787 had 99.1%
  • strain C and YIT 12788 had 99.3% homology of 16S rRNA. Both were 99% or more.
  • the homology between strain C and E. desmolans and B. pulllicaecorum was measured by the method of Example 4 (4)
  • the homology between strain C and E. desmolans was 8.8-18.
  • the homology between strain C and B.pullicaecorum was 10.6 to 15.5%. Therefore, the DNA-DNA homology between strain C and E. desmolans and B. pulllicaecorum was both 20% or less.
  • strain C The ability of strain C to produce butyric acid and formic acid was measured by the method of Example 2 (1). As a result, the amount of butyric acid produced by strain C was 13.3 mM, and the amount of formic acid produced was 0.0 mM. Therefore, the strain C in which the homology of YIT 12787 and YIT 12788 with 16S rRNA is 99% or more and the DNA homology with E. desmolans and B. pulllicaecorum is less than 20% is 1.
  • C. leptum subgroup in which 0 ⁇ 10 6 cells are inoculated into 2 mL of acetic acid-free YCG medium and anaerobic culture at 37 ° C. for 72 hours has a butyric acid production of 10 mM or more and a formic acid production of less than 1 mM It was found to be a butyric acid-producing bacterium belonging to
  • Example 6 Manufacture of tablets Various ingredients were mixed according to the formulation shown in Table 7 below, granulated, dried and sized, and then tableted to produce tablets.
  • a live cell of YIT 12787 was produced by freeze-drying (including 10 10 cells / g of live cell).

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Abstract

L'invention concerne une nouvelle bactérie productrice d'acide butyrique qui appartient au sous-groupe Clostridium leptum et qui, dans un milieu de culture exempt d'acide butyrique, produit de l'acide butyrique mais ne produit pas d'acide formique.
PCT/JP2016/074497 2015-08-24 2016-08-23 Bactérie productrice d'acide butyrique WO2017033925A1 (fr)

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Cited By (3)

* Cited by examiner, † Cited by third party
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JP2020535826A (ja) * 2017-10-02 2020-12-10 メタボリック エクスプローラー 発酵ブロスから有機酸塩を生産する方法
JP2020536865A (ja) * 2017-10-03 2020-12-17 セレス セラピューティクス インコーポレイテッド トリプタミン代謝の操作
CN114250184A (zh) * 2021-12-31 2022-03-29 广东海大集团股份有限公司 一种丁酸梭菌的发酵培养基及发酵工艺

Citations (6)

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