WO2019230183A1 - 乳酸菌及びその用途 - Google Patents
乳酸菌及びその用途 Download PDFInfo
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- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
Definitions
- the present invention relates to a novel lactic acid bacterium or a treated product thereof, a method for growing the lactic acid bacterium or a treated product thereof, and use thereof.
- Lactic acid bacteria are widely used in fermented foods such as yogurt and pickles, and are very familiar and safe. In recent years, some lactic acid bacteria have attracted attention as probiotics, that is, microorganisms that have a positive effect on human health, and researches on the various effects of such lactic acid bacteria are progressing daily.
- lactic acid bacteria having an IgA production inducing action have been reported as one of the functions of lactic acid bacteria (Patent Documents 1 and 2).
- lactic acid bacteria or treated products thereof have an IgA production-inducing action
- lactic acid bacteria or treated products thereof are expected to have effects such as infection prevention, tumor suppression, allergy suppression and intestinal regulation, and the like.
- the body or a processed product thereof can be used as an immunostimulant or an intestinal regulating agent (Patent Document 3).
- the effect brought about by lactic acid bacteria or the processed product thereof may be greatly different if the strains are different even if they are the same bacterial species. Therefore, the selection of strains is very important, and attempts are being made every day to isolate lactic acid strains with higher functions.
- This invention makes it a subject to provide the novel and outstanding lactic acid bacteria or its processed material. Moreover, this invention makes it a subject to provide the manufacturing method of the said lactic acid bacteria or its processed material, or its use.
- E. durans lactic acid bacteria Enterococcus durance
- NITE BP-02675 accession number NITE BP-02675
- this invention includes the following each invention in order to solve the said subject.
- Enterococcus durans HS-08 Enterion number NITE BP-02675 or a treated product thereof.
- An IgA production-inducing or immunostimulating composition containing the bacterial cell according to [1] or a processed product thereof.
- An antiallergic composition containing the bacterial cell according to [1] or a processed product thereof.
- a composition for protecting mucosa comprising the bacterial cell according to [1] or a processed product thereof.
- a plant fertilizer comprising the fungus body according to [1] or a processed product thereof, or the composition according to [7].
- An animal feed comprising the bacterial cell according to [1] or a processed product thereof, or the composition according to [8].
- a food / beverage product, cosmetic or pharmaceutical comprising the microbial cell according to [1] or a processed product thereof, or the composition according to any one of [2] to [8].
- a concrete member comprising the fungus body according to [1] or a treated product thereof, or the composition according to any one of [2] to [8].
- a method of growing Enterococcus durans HS-08 cells which comprises culturing the cells of Enterococcus durance HS-08 (Accession number NITE BP-02675).
- the present invention relates to lactic acid bacteria Enterococcus durans (hereinafter also referred to as “E. durans”) HS-08 strain (Accession No. NITE BP-02675) (hereinafter also referred to as “Lactic acid bacteria of the present invention”).
- E. durans lactic acid bacteria Enterococcus durans
- Lactic acid bacteria of the present invention Provide processed material.
- the lactic acid bacterium of the present invention promotes a remarkable production-inducing action of IgA, which is one kind of immunizing antibody, and an immune activation action in the living body thereby.
- the lactic acid bacterium of the present invention has an antiallergic effect, and more surprisingly, an inhibitory action of harmful bacteria and / or pathogenic bacteria, a mucosal protective action, an action of enhancing short chain fatty acid production, an action of enhancing organic acid production, It exhibits a gene expression increasing effect, plant growth control, lodging prevention, umami control effect, animal growth promotion, or autoimmunity improving effect.
- FIG. 1 is a graph showing the amount of IgA ( ⁇ g / day) in mouse feces.
- FIG. 2 is a graph showing IgA concentration ( ⁇ g / mL) in mouse blood.
- FIG. 3 is a graph showing mouse body weight (g).
- FIG. 4 is a graph showing the CRE concentration ( ⁇ g / mL) in mouse blood.
- FIG. 5 is a graph showing the ALT concentration (iU / L) in mouse blood.
- FIG. 6 is a graph showing the AST concentration (iU / L) in mouse blood.
- FIG. 7 is a graph showing the expression levels of various genes involved in IgA production.
- FIG. 1 is a graph showing the amount of IgA ( ⁇ g / day) in mouse feces.
- FIG. 2 is a graph showing IgA concentration ( ⁇ g / mL) in mouse blood.
- FIG. 3 is a graph showing mouse body weight (g).
- FIG. 4 is a graph showing
- FIG. 8 is a graph showing organic acid and short-chain fatty acid concentrations ( ⁇ mol / g) in mouse feces.
- FIG. 9 is a graph showing the pH in the feces of mice.
- FIG. 10 is a graph showing the gene expression level of GRP43.
- E. durans HS-08 (accession number NITE BP-02675) is a product of the National Institute of Technology and Evaluation Biotechnology Center, Patent Microorganism Depositary Center (NPMD) (address: postal 292-0818, Kazusa, Kisarazu, Chiba, Japan) 2-5-8) was received for international deposit under the Budapest Treaty on April 10, 2018.
- NPMD Patent Microorganism Depositary Center
- the bacteria can be obtained by applying to the deposit center, for example, morphological characteristics (eg, colony shape, cell shape, etc.), physiology, biochemical properties (eg, sugar utilization, growth) Temperature, optimal pH, etc.), chemical taxonomic properties (cell fatty acid composition, etc.) and other properties are compared with E. durans HS-08 (accession number NITE BP-02675) and based on the comparison results
- morphological characteristics eg, colony shape, cell shape, etc.
- biochemical properties eg, sugar utilization, growth
- chemical taxonomic properties cell fatty acid composition, etc.
- the identified bacterium may be used, or the bacterium identified based on the analysis of the base sequence of the 16S rRNA gene.
- the E. durans HS-08 strain is preferably cultured, for example, by the following method.
- Culture medium medium: MRS (de Man, Rogosa and Sharpe) medium, MRS agar medium, LBS medium, modified LBS agar medium, Rogosa medium, other known lactic acid bacteria medium or culture medium, and the like.
- the pH is about 4 to 9, preferably about 5 to 8, and more preferably about 5.5 to 7.5.
- the culture method may be stationary culture, but may also be stirring culture. In the case of stirring culture, the number of stirring may be about 100 to 250 times per minute, preferably about 120 to 180 times per minute, and more preferably about 140 to 160 times per minute.
- the culture time is usually about 1 to 72 hours, preferably about 3 to 48 hours, and more preferably about 6 to 24 hours.
- the culture temperature is usually about 20 to 40 ° C., preferably about 25 to 39 ° C., more preferably about 30 to 38 ° C.
- the MRS medium, MRS agar medium, LBS medium, modified LBS agar medium, Rogosa medium, and other known lactic acid bacteria culture media or culture solutions may contain other additives.
- components or additives contained in such a medium include, but are not limited to, yeast-derived components, soybean-derived components, corn-derived components, animal and plant proteins or extracts thereof, and decomposition products thereof, ammonium nitrate, ammonium sulfate, ammonium chloride, One or more nitrogen sources selected from the group consisting of ammonium salts such as ammonium acetate, ammonia, sodium nitrate, potassium nitrate, sodium glutamate, urea, amino acids, gluten, casein, peptone, glucose, inositol, maltose, xylose, mannose , Fructose, starch, lactose, glycerol, arabinose, ribose, galactose, fructose, inositol, mannitol, sorbitol, glucosamine, cellobiose, sucrose, tre Loin, xylitol, alcohol, starch syrup, starch
- vitamins such as vitamins A, B, C, D, E, and K or their derivatives, or salts thereof, minerals such as zinc, iron, magnesium, potassium, calcium, and phosphorus, pH buffering agents, surfactants Antibiotics, stabilizers and the like, but are not limited thereto.
- Other components or additives include, but are not limited to, sucrose, carbonate, bicarbonate, HEPES, albumin, insulin, amino acids, cytokines, growth factors, hormones and the like.
- An additive can be used individually by 1 type or in combination of 2 or more types. In addition, about anaerobic or aerobic conditions at the time of culture
- the lactic acid bacteria of the present invention may be used in the form of powder by freeze-drying, low-temperature drying, spray drying, L-drying, etc., or a combination thereof, or as obtained by the above method (paste form, liquid form) It may be used as such.
- the viable cell count is preferably 10 7 to 10 8 cfu / g, more preferably 10 9 to 10 12 cfu / g, but is not limited thereto.
- the number of viable bacteria is preferably 10 7 to 10 8 cfu / mL, more preferably 10 9 to 10 12 cfu / mL, but is not limited thereto.
- the measurement of the number of viable bacteria varies depending on the bacterial cells, but can be easily measured by, for example, each bacterial cell quantification method described in the Japanese Pharmacopoeia Pharmaceutical Standards.
- the “treated product” of lactic acid bacteria or the “treated product” of other lactic acid bacteria of the present invention is preferably a processed product of the lactic acid bacteria or a culture of the lactic acid bacteria, but is not limited thereto.
- the bacteria may be live or dead.
- such a treated product may be used as it is, or may be used in the form of powder by freeze drying, low temperature drying, spray drying, L-drying, or the like, or a combination thereof.
- These processed products (cultured products) may be used after diluted with an appropriate solvent (water, alcohol, organic solvent, etc.), or added to an appropriate additive to form a gel or solid agent. May be.
- the lactic acid bacterium of the present invention or a processed product thereof is used in combination with other useful bacteria (for example, a lactic acid bacterium belonging to the genus Lactobacillus, Leuconostoc, Streptococcus; bifidobacteria; yeast, or mold) or a processed product thereof. May be.
- other lactic acid bacteria include Lactobacillus acidophillus, Lactobacillus delbrueckii, Lactobacillus casei, Lactobacillus reichmannii, Lactobacillus Fructivorans (L.) fructivorans), Lactobacillus helveticus (L. helveticus), Lactobacillus hilgardii (L.
- Lactobacillus kefiri (L. kefiri), Lactobacillus kefiranofaciens (L. kefiranofaciens), Lactobacillus kefirgranum (L. kefirgranum), Lactobacillus parakefia (L. parakefir), Lactobacillus pentoaceticus (L. pentoaceticus), Lactobacillus cullet (L. caret), Lactobacillus bulgaricus (L . Bulgaricus), lact L. lactis, L. plantarum, Lactobacillus paracasei, L. cellobiosus, Lactobacillus confusus (L.
- Lactobacillus confusus Lact Bacillus coprophilus (L. coprophilus), Lactobacillus fermentum (L. fermentum), Lactobacillus San Francisco (L. sanfrancisco), Lactobacillus thermophilus (L. thermophilus), Lactobacillus bavaricus (L. bavaricus), Lactobacillus brevis (L. brevis), Lactobacillus buchneri (L. buchneri), Lactobacillus coriniformis (L. coryniformis), Lactobacillus carvatus (L. curvatus), Lactobacillus salmon (L.
- Lactobacillus Lactobacillus, Leuconostoc Cremo Leuconostoc cremoris, Leuconostoc genus such as Leuc.
- Examples of the genus Streptococcus include, but are not limited to. These useful bacteria can be cultivated by a conventionally known method, and from, for example, an organization such as ATCC (registered trademark) or IFO, the Japan Bifidobacteria Center, an independent administrative agency, the Product Evaluation Technology Infrastructure Organization Patent Microorganism Deposit Center, etc. It can be easily obtained. Moreover, what is marketed can also be used suitably.
- Examples of bifidobacteria that can be used in combination with the lactic acid bacterium of the present invention or a processed product thereof include, for example, Bifidobacterium bifidum, B. angulatum, Bifidobacterium adduct Lessentis (B. adolescentis), Bifidobacterium breve (B. breve), B. ⁇ ⁇ catenulatum, B. dentium (B. dentium), Bifidobacterium B. ⁇ ⁇ gallicum, B.subtile, B. asteroides, B. boum, Bifidobacterium coelinum ( B. choerinum), B.
- Bifidobacte Um Kunikri (B. cuniculi), Bifidobacterium gallinarum (B. gallinarum), Bifidobacterium indicum (B. indicum), Bifidobacterium magnum (B. magnum), Bifidobacterium ⁇ Mericicum (B. merycicum), Bifidobacterium minimum (B. minimum), Bifidobacterium psychraerophilum (B. psychraerophilum), Bifidobacterium prolum (B. pullorum), Bifidobacterium ⁇ B. ruminantium, B. ⁇ scardovii, B. thermophilum, B.
- infantis Bifidobacterium ⁇ Mongolianse (B. ⁇ mongoliense), Bifidobacterium pseudocatenulator (B. pseudocatenulatum), B. ⁇ thermacidophilum subsp. Porcinum, B. ⁇ animalis subsp. Lactis, Bifidobacterium. Animalis subspecies animalis (B. animalis subsp. Animalis), Bifidobacterium ⁇ ⁇ ⁇ ⁇ longum subspecies longum (B. longum subsp. Longum), Bifidobacterium subspecies pseudolongum (B. pseudolongum subsp.
- Bifidobacterium examples include, but are not limited to, the genus Bifidobacterium such as B. pseudolongum subsp. Globosum or its subspecies. These bifidobacteria can be cultured by a conventionally known method, and can be easily obtained from various institutions and centers similar to the above-mentioned lactic acid bacteria, for example. Moreover, what is marketed can also be used suitably.
- yeast examples include Saccharomyces cerevisiae (also known as budding yeast, top fermenting yeast), Saccharomyces kefir (S. kefir), Saccharomyces lactis (S. lactis), Saccharomyces Karlsbergensis (S. carlsbergensis (also known as bottom fermentation yeast)), Saccharomyces unisporus, S. pastrianus, Saccharomyces bayanus (S. bayanus), Saccharomyces Karlsbergensis (S. carlsbergensis), Saccharomyces delbrueckii (S.
- Saccharomyces dirensis S. dairensis
- Saccharomyces diatatics S. diataticus
- Saccharomyces et Saccharomyces genus such as Goose (S. exiguus), Saccharomyces kluyveri (S. Vietnamese)
- Saccharomyces lucy S. Pombe (Schizosaccharomyces pombe (also known as fission yeast)), Schizosaccharomyces rouxii, etc.
- Candida utilis also known as Torula yeast
- Candida tropica Squirrel C. tropicalis
- Candida Milleri C. milleri
- Candida Crusei C.
- Candida Lucitaniae C. lusitaniae
- Candida Aaseri C. guilliermondii (C. guilliermondii) ), C. humicola, C. glabrata, C. lambica, C. lipolytica, C. parapsilosis, C. tropicalis, Candida paratropica Candida genus such as squirrel (C. paratropicalis), Candida pseudotropicalis (C. rugosa), Candida stellatoidea (C. stellatoidea), Candida zeylanoides (C. zeylanoides), etc.
- -Torulaspora genus such as Torulaspora delbrueckii, Torulopsis celluculosa, Torulopsis genus such as Torulopsis candida, Torula genus such as Torula kefir, Cerberus cerevisias (Kluyveromyces bulgaricus) Cerberymyces fragilis (K. fragilis), Cerberymyces thermotolerans (K. thermotolerans), Cerberymyces lactis (K. lactis), Cerberymyces marxianus (K. marxianus), Cerberymyces fragilis (K. fragilis) and other genus Kerberomyces, P. membranaefaciens, P.
- yeasts can be cultured by a conventionally known method, and can be easily obtained from various institutions and centers similar to the above lactic acid bacteria. Moreover, what is marketed can also be used suitably.
- Examples of other useful bacteria that can be used in combination with the lactic acid bacterium of the present invention or a processed product thereof include Acetobacter aceti, Acetobacter orientalis, Acetobacter pasteurianus, Acetobacter pasteurianus, Acetobacter pasteurianus ⁇ Acetic acid bacteria including Acetobacter genus such as Acetobacter xylinum, Butyric acid bacteria including Clostridium butyricum, Pediococcus damnosus, Pediococcus ospentosaceus, Pediococcus genus such as Pediococcus halophilus, filamentous fungi (Bacillus), Bacillus subtilis natto, Bacillus subtilis, actinomycetes, photosynthetic bacteria, amino acid-producing bacteria, cellulose-degrading bacteria ( For example, Clostridium , Trichoderma, etc.), Rhizobium, mycorrhizal fungi, VA bacteria, but such nitrifying bacteria
- molds that can be used in combination with the lactic acid bacteria of the present invention or processed products thereof include Aspergillus oryzae, Aspergillus sojae, Aspergillus mucor, Aspergillus niger, Aspergillus niger, Aspergillus flavus, Aspergillus nidulans and other Aspergillus genus, Penicillium roqueforti, Penicillium ⁇ Rhizopus and Penicillium ⁇ Rhizopus, etc. Not. These molds can be cultured by a conventionally known method, and can be easily obtained from, for example, various institutions and centers similar to the above-mentioned lactic acid bacteria. Moreover, what is marketed can also be used suitably.
- the lactic acid bacterium of the present invention or a processed product thereof, or a composition containing the lactic acid bacterium may be directly sprayed on plants (leaves, stems, fruits, etc.) or sprayed (administered) on soil. Alternatively, it may be mixed with plant fertilizer, or in the case of field cropping or rice cropping, it may be installed at a water intake or the like, or may be mixed as a concrete raw material of the water intake or a material of a concrete structure. Moreover, when the lactic acid bacteria of this invention, its processed material, or the composition containing this is applied to the livestock field, it may mix and give to animal feed or drinking water, a wall, a floor, etc. in cowshed, a poultry house, etc. Alternatively, it may be mixed as a concrete raw material or a concrete structure material.
- the present invention provides a composition containing the lactic acid bacteria of the present invention.
- known additives usually used in the art can be used.
- water, solvent, pH adjuster, humectant, flavor, sweetener, thickener, flavoring agent, Gelling agents, solubilizers, colorants, preservatives, surfactants, suspending agents, emulsifiers, stabilizers and the like can be mentioned, but not limited thereto.
- the pH adjuster that can be used in the present invention may be any agent that adjusts the pH of the composition of the present invention to a desired value, and has a pH of about 2.5 to 10.0, more preferably about pH. It can be used to adjust to the range of 5.0 to 8.5.
- Preferred pH adjusters include alkali metal hydroxides, alkaline earth metal hydroxides, alkaline earth metal oxides, alkali metal oxides, carbonates, borates, silicates, phosphates, organic acids , And organic bases.
- phosphoric acid or a salt thereof benzoic acid or a salt thereof, salicylic acid or a salt thereof, sodium hydroxide, potassium hydroxide, sodium carbonate, potassium carbonate, magnesium carbonate or other alkali metal carbonate, magnesium hydroxide, water Alkaline earth metal carbonates such as calcium oxide, alkali metal bicarbonates such as potassium hydrogen carbonate and sodium hydrogen carbonate, alkaline earth metal bicarbonates such as magnesium hydrogen carbonate and calcium hydrogen carbonate, boric acid, citric acid, fumarate Examples include, but are not limited to, acid salts, malic acid, tartaric acid, succinic acid, maleic acid or salts thereof, lactic acid, imidazole, triethanolamine, diethanolamine, trometamol, meglumine, lidocaine, and salts thereof.
- Preferred moisturizers that can be used in the present invention include, but are not limited to, polyhydric alcohols such as sorbitol, xylitol, glycerin, butylene glycol, polyethylene glycol and propylene glycol.
- polyhydric alcohols such as sorbitol, xylitol, glycerin, butylene glycol, polyethylene glycol and propylene glycol.
- flavoring agents examples include peppermint oil, spearmint oil, mint oil, menthol, anethole, sage, lemon oil, orange oil, cinnamon, vanillin, thymol and linalool. It is not limited.
- Sweetening agents that can be used in the present invention include, for example, sucrose, glucose, saccharin, glycyrrhizic acid, dextrose, fructose, lactose, mannitol, sorbitol, fructose, maltose, xylitol, honey, chickenpox, saccharin, aspartame, D- Examples include, but are not limited to, tryptophan, acesulfame, cyclamic acid, and salts thereof.
- flavoring agent examples include ascorbic acid, citric acid, glycyrrhizic acid, glutamic acid, succinic acid, tartaric acid, fumaric acid, malic acid, taurine, sarcosine, glycyrrhizic acid or a salt thereof, or erythritol, Lactitol, reduced palatinose, sodium chloride, magnesium chloride, orange oil, saffron oil, salamander oil, perilla oil, basil oil, peppermint oil, lemon oil, lemongrass oil, rose oil, rosemary oil, cacao, caramel, licorice, Camphor, cinnamon oil, saffron, methyl salicylate, peony extract, ginger, cinnamaldehyde, stevia extract, assembly, sorbitol, cyclodextrin, soybean oil, tisoiso extract, taurine, tannic acid, clove oil, spruce , Nigakiekisu
- Surfactants that can be used in the present invention may be nonionic, zwitterionic, anionic, or cationic. Any nonionic surfactant can be used, for example, polyoxyethylene sorbitan fatty acid ester such as polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 80; polyoxyethylene hydrogenated castor oil 10, polyoxyethylene hydrogenated castor Polyoxyethylene hydrogenated castor oil such as oil 40, polyoxyethylene hydrogenated castor oil 50, polyoxyethylene hydrogenated castor oil 60; polyoxyethylene polyoxypropylene glycol; polyethylene glycol monolaurate, ethylene glycol monostearate, polyoxyl stearate 40 Polyethylene glycol fat such as polyethylene glycol monostearate, polyethylene glycol monooleate, ethylene glycol monostearate Ester, and the like, but not limited thereto.
- anionic surfactant examples include sodium alkylbenzene sulfonate and ammonium dodecylbenzene sulfonate, but are not limited thereto.
- zwitterionic surfactant examples include aliphatic derivatives of secondary or tertiary amines containing carboxy, sulfonate, sulfate, or aliphatic secondary derivatives of heterocyclic secondary or tertiary amines, etc. Examples thereof include, but are not limited to, (C 8-20 alkyl) betaine and (C 8-20 alkyl) amide (C 6-8 alkyl) betaine, and mixtures thereof.
- Cationic surfactants include, but are not limited to, quaternary ammonium salts, amine salts, amines, and the like.
- the emulsifier and suspending agent that can be used in the present invention, the above-mentioned surfactants can be suitably used.
- lecithins such as soybean lecithin, egg yolk lecithin, hydrogenated lecithin, enzymatically decomposed lecithin, Higher alcohols such as cetanol, lauryl alcohol, stearyl alcohol, and lanolin alcohol can be used, but are not limited thereto.
- Stabilizers that can be used in the present invention include sodium polyacrylate, adipic acid, ascorbic acid, sodium sulfite, sodium bisulfite, dibutylhydroxytoluene, butylhydroxyanisole, sodium edetate, sodium chloride, citric acid, cyclodextrin , Cysteine and the like, but are not limited thereto.
- the lactic acid bacterium of the present invention or a processed product thereof, or a composition containing the lactic acid bacterium may be administered to animals including humans.
- the lactic acid bacterium of the present invention or a processed product thereof, or a composition containing the same may be administered as feed (agricultural materials), medicine, food, etc. itself or as a raw material thereof, and may be administered, pharmaceuticals, cosmetics, etc. It may be applied as such or as its raw material.
- the animal to which the composition of the present invention is administered is not particularly limited, and may be, for example, a human or a non-human animal.
- animals other than humans include, but are not limited to, for example, domestic animals such as cattle, horses, pigs, and sheep, dogs, cats, rabbits, hamsters, squirrels, guinea pigs, mice, and other pets or laboratory animals, sushi, tuna, Shark, sunfish, ray, horse mackerel, sea bream, sea bream, sea bream, sea bream, sea bream, sea bream, sea bream, sea bream, fish, fish, shrimp, sea bream, sea bream, scallop, oyster, Fish and shellfish, chickens, quail, turkeys, ducks, geese, parakeets, parrots and other birds, beetles, stag beetles, swallowtail butterflies, praying mantis, grasshoppers, grasshoppers, bellworms, damselfly, geckos, chameleons, iguanas, lizards Examples include reptiles, amphibians such as newts, f
- the administration form of the present invention is not particularly limited, and examples thereof include oral administration and parenteral administration (intravenous administration, transdermal administration, etc.).
- Examples of the dosage form of the present invention include tablets, capsules, granules and powders in the case of oral preparations, and examples of parenterals include injections, ointments and insertion agents.
- the dosage form of the present invention is, for example, a tablet, capsule, granule, powder or the like, it is not particularly limited.
- excipients such as mannitol, calcium phosphate, starch, sucrose, lactose, glucose
- Disintegrants such as methylcellulose, starch, pregelatinized starch, carboxymethylcellulose calcium, croscarmellose sodium, crospovidone, low substituted hydroxypropylcellulose; hydroxypropylcellulose, ethylcellulose, gum arabic, starch, partially pregelatinized starch, polyvinylpyrrolidone, Binders such as polyvinyl alcohol; lubricants such as magnesium stearate, calcium stearate, talc, hydrous silicon dioxide, hydrogenated oil; polyvinyl pyrrolidone, sugar, hydroxypropylmethyl Loin, hydroxypropyl cellulose, methyl cellulose, and suitably selected according to need, such as coating agents such as ethyl
- the dosage form when the dosage form is an injection, it is not limited to, but is not limited to, for example, an isotonic agent such as sodium chloride; a buffer such as sodium phosphate; A surfactant such as oxyethylene sorbitan monooleate; a thickener such as methylcellulose, and the like can be appropriately selected and used as necessary.
- an isotonic agent such as sodium chloride
- a buffer such as sodium phosphate
- a surfactant such as oxyethylene sorbitan monooleate
- a thickener such as methylcellulose, and the like
- the preparation of the present invention is locally administered to the eye, for example, when the dosage form is an eye drop or the like, but is not limited thereto, for example, isotonic agents such as sodium chloride and concentrated glycerin; sodium phosphate, acetic acid Buffering agents such as sodium; surfactants such as polyoxyethylene sorbitan monooleate, polyoxyl 40 stearate, polyoxyethylene hydrogenated castor oil; stabilizers such as sodium citrate and sodium edetate; benzalkonium chloride, parabens
- a preservative such as, for example, can be appropriately selected and used as necessary.
- the pH adjuster for example, those described below can be used.
- the preparation of the present invention is an ointment, transdermal agent, patch, etc.
- petrolatum for example, petrolatum, squalane, paraffin, liquid paraffin, lauric acid, myristic acid, stearic acid, isostearic acid, oleic acid and other higher fatty acids, beeswax
- a general-purpose base such as oils and fats such as waxes such as lanolin.
- the preparation of the present invention is an intercalating agent
- it can be formulated using, for example, a biodegradable polymer such as hydroxypropylcellulose, hydroxypropylmethylcellulose, carboxyvinyl polymer, polyacrylic acid, and the like.
- a biodegradable polymer such as hydroxypropylcellulose, hydroxypropylmethylcellulose, carboxyvinyl polymer, polyacrylic acid, and the like.
- the dose of the present invention can be appropriately selected according to the dosage form, patient's symptoms, age, weight, etc., but is not particularly limited.
- 0.05 to 5000 mg, preferably 0.1 to 2000 mg, particularly preferably 1 to 1000 mg per day per kg of body weight can be administered in 1 to several times a day.
- 0.0001 to 2000 mg per kg body weight per day, preferably 0.001 to 1500 mg, particularly preferably 0.01 to 500 mg can be administered in 1 to several divided doses per day.
- This invention provides the foodstuff containing the lactic acid bacteria of this invention, its processed material, or the composition containing this.
- the food includes health food, functional label food, food for specified health use, and food for the sick.
- the form of the food is not particularly limited. Specific examples include, but are not limited to, tablets, granules, powders, drinks and the like as so-called dietary supplements or supplements.
- beverages such as soft drinks, nutritional drinks, carbonated drinks, fruit drinks, lactic acid drinks, tea drinks, noodles such as buckwheat, udon, Chinese noodles, instant noodles, chocolate, snacks, candy, gum, biscuits , Jelly, jam, cream and other confectionery, breads, sausages, kamaboko, ham and other processed marine products, processed milk, fermented milk (including yogurt) and other dairy products, margarine, mayonnaise, shortening, whipped cream, dressing Fats such as salad oil and tempura oil or processed foods, soy sauce, seasonings such as sauces, retort pouch foods such as curry, rice cakes, rice cakes, miscellaneous foods, stews, and frozen desserts such as ice cream and sherbet .
- the proportion of E. durans HS-08 strain in the food is preferably 0.1 to 10%, more preferably 1 to 10%, based on the total weight of the food.
- the present invention provides a food additive containing the lactic acid bacterium of the present invention or a processed product thereof, or a composition containing the same.
- food additives include, but are not limited to, pH adjusters, preservatives, bactericides, antioxidants, fungicides, coloring agents, coloring agents, bleaching agents, brighteners, fragrances, spices extraction Products, softeners, nutrient enhancers, sweeteners, acidulants, seasonings, bitters, emulsifiers, thickeners, stabilizers, gelling agents, pastes, swelling agents, manufacturing solvents, enzymes, gum bases, yeast foods These preparations can be mentioned.
- the additives exemplified above can also be used in the same manner.
- the proportion of E. durans HS-08 in the food additive is preferably 0.1 to 10%, more preferably 1 to 10%, based on the total mass of the food additive.
- Cosmetics include so-called medicinal cosmetics (quasi-drugs). Cosmetics include, but are not limited to, for example, detergents, shampoos, rinses, hair nicks, hair lotions, after shave lotions, body lotions, cosmetic lotions, cleansing creams, massage creams, emollient creams, aerosol products, deodorants, A fragrance
- the cosmetics of the present invention are components commonly used as cosmetics other than the lecithin of the present invention, for example, surfactants, moisturizers, animal and plant-derived oils and fats, microorganism-derived oils and fats, silicones, higher alcohols, lower alcohols, animal and plant-derived extracts. Extracts, microorganism-derived extracts, UV absorbers, anti-inflammatory agents, sequestering agents, vitamins, antioxidants, thickeners, preservatives, bactericides, pH adjusters, colorants, various flavors, etc. It can mix
- the proportion of E. durans HS-08 in the cosmetic is preferably 0.1 to 10%, more preferably 1 to 10%, based on the total mass of the cosmetic.
- This invention provides the pharmaceutical containing the lactic acid bacteria of this invention, its processed material, or the composition containing this.
- the pharmaceutical preparation can be formulated by containing an active ingredient and further appropriately blending pharmaceutically acceptable carriers and additives as necessary.
- Pharmaceuticals are not limited to these, but specifically, tablets, coated tablets, pills, powders, granules, capsules, solutions, suspensions, emulsions, injections, infusions, suppositories, ointments, sprays, Ointments, creams, gels, patches, and the like can be used.
- Carriers or additives that can be blended are not particularly limited.
- various carriers such as water, physiological saline, other aqueous solvents, aqueous or oily bases, excipients, binders, pH adjusters, disintegrants, Various additives such as absorption accelerators, lubricants, colorants, flavoring agents, and fragrances can be mentioned. In addition to these, those exemplified above can also be preferably used.
- the proportion of E. durans HS-08 strain in the pharmaceutical is preferably 0.1 to 10%, more preferably 1 to 10%, based on the whole pharmaceutical.
- This invention provides the animal feed containing the lactic acid bacteria of this invention, its processed material, or the composition containing this.
- feed include feed for livestock such as cattle, horses, pigs and sheep, feed for pets such as dogs, cats, rabbits, hamsters, squirrels, guinea pigs and rats, sardines, tuna, sharks and sunfish.
- the feed of the present invention can be produced using a general feed production method, except that the lactic acid bacteria of the present invention are added to the feed.
- the ratio of E. durans HS-08 strain in the feed is preferably 0.1 to 10% by weight, more preferably 1 to 10% by weight, based on the total feed.
- the animal feed leads to promotion of animal growth or improvement of autoimmunity, shortening of the breeding period of animals fed with the feed (particularly livestock or poultry), or early shipment.
- This invention provides the fertilizer for plants containing the lactic acid bacteria of this invention, its processed material, or the composition containing this.
- the fertilizer include, but are not limited to, vegetables, flowers, flowering trees, fruits and the like.
- root crops such as lotus root, potatoes, sweet potatoes, potatoes, taro, taro, konjac potatoes, yams, edible beans, empty beans, peas, Beans such as green beans, lentils, red beans, sesame seeds, cabbage, lettuce, Chinese cabbage, spinach, Nozawana, mizuna, trefoil, komatsuna, spring chrysanthemum leaf vegetables, tangerine, sweet summer, bundan, tangerine, Iyokan, Yawata, lemon Citrus fruits such as lime, grapefruit, onions, long onions, white onions, cucumbers, enoki, garlic, jellyfish, shimeji, nameko, matsutake mushrooms, shiitake mushrooms, maiko, broccoli, cauliflower, rapeseed, Flowers and vegetables such as artichokes, cucumbers, watermelons, pumpkins, zucchini, gourds, loofahs, to
- the fertilizer of this invention can be manufactured using the manufacturing method of a general fertilizer except adding the lactic acid bacteria of this invention in a fertilizer.
- the proportion of E. durans HS-08 strain in the fertilizer is preferably 0.1 to 10% by mass, more preferably 1 to 10% by mass, based on the entire fertilizer.
- the said fertilizer for plants may have effects such as plant growth regulation, plant lodging prevention, or plant taste adjustment.
- the plant fertilizer can, for example, promote the growth of plant roots, stems, branches, leaves, fruits, or flowers.
- the said fertilizer for plants can enhance the umami
- the present invention provides an industrial product (for example, a concrete member) containing the lactic acid bacterium of the present invention or a processed product thereof, or a composition containing the lactic acid bacterium.
- industrial products other than concrete members include, but are not limited to, petroleum products, resin products, magnetic products, leather products, and fabrics.
- the proportion of E. durans HS-08 in the industrial product is preferably 0.1 to 10% by mass, more preferably 1 to 10% by mass, based on the industrial product.
- the industrial product is the lactic acid bacterium of the present invention or a processed product thereof, which can have effects such as promotion of animal growth, improvement of animal autoimmunity, plant growth regulation, plant lodging prevention, or plant taste control.
- Industrial products used for animal breeding for example, bait boxes, fences, etc.
- industrial products used for plant growth for example, potted plants, watering cans, buckets, hoses, concrete for intakes, etc.
- the kit containing the lactic acid bacteria of this invention, its processed material, or the composition containing this is provided.
- the kit preferably consists of a container that can contain the composition separately, such as, for example, a split bottle or foil packet, but the composition can also be contained in a single container.
- a container that can contain the composition separately, such as, for example, a split bottle or foil packet, but the composition can also be contained in a single container.
- preferable kits include, but are not limited to, blister packs used for packaging tablets and capsules, syringes and containers filled with a drug solution, and the like.
- the lactic acid bacteria of the present invention preferably have an immunostimulatory effect.
- the immunostimulatory action for example, when the administration group of the lactic acid bacterium or the lactic acid bacterium-containing composition of the present invention shows a significant difference or significant tendency in the fecal or blood IgA concentration compared to the non-administration group, It can be determined that it has an effect.
- symptoms of immune diseases eg, mastitis, colibacillosis, diarrhea, etc.
- This can also reduce the amount of antibiotic used.
- IgA produced on Peyer's patches prevents such adhesion and plays an important role in the immune system.
- the Peyer's patch is in the intestinal tract and is composed of T cells, B cells, plasma cells that produce IgA, and the like.
- acetic acid is known to induce IgA by inducing retinoic acid.
- IL-6, RALDH2, APRIL secreted by dendritic cells of Peyer's patch , BAFF, etc. are also known to induce IgA production.
- the lactic acid bacteria or lactic acid bacteria-containing composition administration group of the present invention is significantly different or significantly different from the non-administration group in acetic acid concentration and RALDH2, IL-6, APRIL, BAFF gene expression levels.
- the mucosal tissue is preferably a digestive organ, a respiratory organ, a urinary organ, or the like, and more preferably, the immune system of the mucosal tissue of the intestinal tract is activated.
- the lactic acid bacteria of the present invention preferably have a mucosal protective action.
- Organic acids such as lactic acid and acetic acid produced by intestinal bacteria contribute to various biological reactions in animals including humans, but such organic acids are absorbed from mucosal tissues and become an energy source for mucosal epithelial cells, It is also known to repair epithelial cells and protect mucosa. Therefore, for example, when the administration group of the lactic acid bacteria or lactic acid bacteria-containing composition of the present invention shows a significant difference or significant tendency in the amount of lactic acid or acetic acid in the mucosal tissue compared to the non-administration group, the mucosal protective effect is exhibited. It can be judged that it has.
- the lactic acid bacteria of the present invention preferably have an antiallergic action.
- the administration group of the composition containing lactic acid bacteria of the present invention has a significant difference or significant tendency in IgA concentration in blood or feces compared to the non-administration group, it is judged to have an antiallergic effect. sell.
- symptoms such as hay fever can be alleviated.
- the lactic acid bacterium of the present invention preferably has an effect of enhancing production of short chain fatty acids or organic acids. It is known that short chain fatty acids or organic acids produced by intestinal bacteria such as lactic acid bacteria contribute to the production of IgA. Therefore, for example, the administration group of the lactic acid bacteria or lactic acid bacteria-containing composition of the present invention is not administered. If the concentration of the short chain fatty acid or organic acid is high with a significant difference or significant tendency compared to the group, it can be determined that the production of the short chain fatty acid or organic acid is enhanced.
- Examples of the short-chain fatty acid or organic acid that the lactic acid bacterium of the present invention enhances production include, but are not limited to, lactic acid and acetic acid.
- Short chain fatty acids are known to bring about an immunostimulatory action and an antiallergic action by IgA, and are considered to have various positive effects on human and animal organisms.
- the lactic acid bacterium of the present invention preferably has an effect of increasing the gene expression of the acetate receptor GRP43.
- GRP43 is a receptor having a high affinity for short-chain fatty acids, particularly acetic acid, and it is known that GRP43 increases as the amount of short-chain fatty acids in the intestine increases. That is, if the gene expression level of GRP43 increases, it is considered that short chain fatty acids (such as acetic acid) in the intestinal tract increase.
- the increase in GPR43 gene expression has various useful effects such as diabetes prevention effect through regulation of insulin signal, colitis suppression effect by induction of regulatory T cells involved in homeostasis.
- the lactic acid bacterium of the present invention preferably has an effect of inhibiting the growth of harmful bacteria and / or pathogenic bacteria. It is known that harmful bacteria and pathogens for animals such as humans and mice are suppressed in an acidic environment, so if the organic acid such as lactic acid or acetic acid increases, the intestinal environment is acidified, It is considered that the pH is lowered and the growth of harmful bacteria and pathogenic bacteria in the intestine is suppressed.
- the administration group of the lactic acid bacterium or lactic acid bacterium-containing composition of the present invention shows a significant difference or a significant tendency in the pH in the intestine compared to the non-administration group, the effect of inhibiting the growth of harmful bacteria and / or pathogens is reduced. It can be judged that it has.
- harmful bacteria and pathogenic bacteria that can suppress growth in the present invention include Clostridium perfringens and Escherichia coli, they are not limited to these.
- the lactic acid bacteria of the present invention preferably have a plant growth regulating action.
- a plant growth regulating action when the administration group of the lactic acid bacteria or lactic acid bacteria-containing composition of the present invention is significantly grown in the size of roots, leaves, branches, fruits or flowers compared to the non-administration group, for example, these sizes are preferable.
- the lactic acid bacteria of the present invention preferably have an action to prevent plant lodging.
- the degree of lodging can be expressed, for example, in six levels from zero (0) to cocoon (5) by the magnitude of the inclination of the plant that has fallen.
- the administration group of the lactic acid bacteria or the lactic acid bacteria-containing composition of the present invention may preferably prevent 1 to 5 stages, more preferably 2 to 4 stages of lodging with respect to the lodging rate of the non-administration group.
- the plant include those exemplified above, and rice or paddy rice is particularly preferable, but is not limited thereto.
- the composition may further contain minerals such as calcium, zinc, iron, magnesium, potassium, and phosphorus. The combined use of minerals can further improve the plant lodging prevention effect. This is thought to be due to the plant's autoimmunity-improving effect, which can reduce the amount of fertilizer used.
- the lactic acid bacterium of the present invention preferably has an effect of adjusting plant umami (sugar content).
- the sugar content may increase by about 1% to 70%, more preferably about 20% to 50%, but is not limited to the original sugar content.
- examples of the plant include those described above, and rice, melon, mandarin orange, lemon, tomato, Chinese cabbage and the like are particularly preferable, but are not limited thereto.
- the lactic acid bacteria of the present invention preferably have an animal growth promoting action.
- the administration group of the lactic acid bacteria or lactic acid bacteria-containing composition of the present invention preferably has an increase in body weight of about 10 to 200% by mass, more preferably about 30 to 100% by mass relative to the body weight of the non-administration group.
- the body length may increase by about 10 to 200%, more preferably about 30 to 100%, but is not limited to the body length of the non-administration group. This can also reduce the amount of feed used.
- animals in the present disclosure include, but are not limited to, those described above.
- the lactic acid bacteria of the present invention preferably have an effect of improving animal autoimmunity.
- the administration group of the lactic acid bacteria or lactic acid bacteria-containing composition of the present invention has an autoimmunity improving action when the stool or blood IgA concentration is significantly different or higher than the non-administration group. Can be judged.
- symptoms of autoimmune diseases eg, hay fever, allergies, etc.
- animals in the present disclosure include, but are not limited to, those described above.
- Example 1 IgA production-inducing activity of E. durans HS-08 strain (1)
- the test mouse is BALB / cA, the age at arrival is 6 weeks, and the sex is male.
- mice immediately after arrival may be affected by stress due to environmental changes and may affect the test results, acclimatization to receive only purified feed (AIN-76A, Research Diets, New Brunswick, NJ) The period was set for 17 days, and thereafter, the period for feeding the feed described below and the observation period were set (about 60 days).
- mice were divided into 4 groups so that body weight and fecal IgA amount were as equal as possible, E.durans HS-08 strain-free classification (hereinafter referred to as control group), E. The ratio of durans08HS-08 shares was 0.38% (hereinafter referred to as 0.38% group), and the number of tests was 5 in each category (Table 1).
- E. durans HS-08 strain was cultured in MRS medium (Difco Laboratories, Detroit, MI, USA) as the lactic acid bacteria powder to be blended in the feed to be ingested by the mice. Specifically, first, a colony of E. durans HS-08 strain was inoculated into 5 mL of MRS culture solution, and statically cultured at 37 degrees in a sealed state for 24 hours to obtain a culture solution. Then, this culture solution was put into 2000 mL of MRS culture solution, and statically cultured at 37 degrees in a sealed state for 24 hours.
- MRS medium Difco Laboratories, Detroit, MI, USA
- the viable cell count of the lactic acid bacteria powder was 1.1 ⁇ 10 12 cfu / g.
- mice On day 0, 14, 28, 42, and 60 after ingesting lactic acid bacteria powder, feces for one day of mice were collected. The feces were mixed with a phosphate physiological saline containing a protease inhibitor and placed on ice for 30 minutes. The mixture was centrifuged, and the IgA concentration of the supernatant was measured using a mouse IgA ELISA kit.
- the 0.38% group has a tendency to increase the IgA amount at the 28th day as compared with the control group, and the IgA amount is significantly increased at the 42nd day. It was. On day 60, the amount of IgA decreased from day 42, but tended to be higher than in the control group.
- the data of the amount of IgA in feces was analyzed by a two-way repeated measurement dispersion method, and then statistical analysis was performed by a multiple comparison test. Other data were statistically analyzed by multiple comparison test.
- the Ig8 concentration in the blood was significantly higher in the 0.38% group than in the control group.
- the group (0.38% group) ingesting the lactic acid bacteria powder had a higher amount of IgA in feces than the control group. This confirmed that E. durans HS-08 can activate the intestinal immune system. Moreover, the group (0.38% group) ingested lactic acid bacteria powder had a higher IgA concentration in the blood than the control group. Since it can be said that the improvement in the IgA concentration in the blood is an improvement in systemic immunity, it was confirmed that the E. durans HS-08 strain can activate the systemic immune system.
- Example 2 The plasma obtained in Example 1 was measured for creatinine (hereinafter, CRE) concentration using lab assay creatinine. Creatinine (CRE) levels in the blood increase when there is an abnormality in the kidney.
- CRE creatinine
- the purpose of this study is to compare the CRE concentrations of the control group and the group ingested with lactic acid bacteria powder, and to confirm the safety of the mouse for renal function.
- ALT alanine aminotransferase
- AST aspartate aminotransferase
- Example 1 the IgE concentration of the plasma obtained in Example 1 was measured using a mouse IgE ELISA kit. The purpose of this test is to compare the IgE concentrations of the control group and the group fed with lactic acid bacteria powder to confirm that the lactic acid bacteria are not allergic to mice.
- Example 3 Genetic analysis of Peyer's patch cells The mice obtained in (1) to (4) of Example 1 were euthanized by cervical dislocation on the 60th day after ingestion of lactic acid bacteria powder. Plate cells were harvested and RNA was extracted using the illustra RNAspin Mini RNA Icolation kit (GE Healthcare, Piscataway, NJ, USA). 0.15 ⁇ g of the RNA was measured and reverse transcribed using an oligo primer (Invitrogen, Carlsbad, Calif.) And Super Script III reverse transcriptase (Invitrogen).
- cDNA was purified from the reverse-transcribed sample using the PCR Purification kit (Qiagen, Cambrige, MA, USA), and gene expression analysis of immune inducing factors in the intestine was performed by real-time PCR using SYBR Green (Life Technologies). went.
- the gene expression levels of RALDH2, APRIL, BAFF, and IL-6 in the group fed with lactic acid bacteria powder tended to be significantly higher or higher than those in the control group.
- IL-6, BAFF, and APRIL secreted from stimulated dendritic cells promote IgA production from B cells
- retinoic acid produced by RALDH2 of dendritic cells is the mucosa of IgA-producing B cells. Promote homing to Therefore, it is clear from the results of FIG. 7 that the lactic acid bacteria of the present invention also enhanced IgA in the mucosa of the intestinal tract at the gene expression level.
- RALDH2 retinal dehydrogenase 2
- APRIL APRIL
- BAFF B-cell activating factor
- IL-6 interleukin-6
- the endogenous control gene used in is ⁇ -actin.
- the primers described in Table 7 below were used as primers for gene analysis.
- Example 4 Short-chain fatty acid or organic acid and pH measurement
- the mice obtained in (1) to (4) of Example 1 were treated with the feces for one day from the mice on the 60th day after feeding the lactic acid bacteria powder. After collection, the feces were lyophilized and suspended in ultrapure water. Thereafter, the mixture was placed at 4 ° C. for 1 hour and centrifuged, and the supernatant was filtered. The filtered supernatant was treated with a short-chain fatty acid analysis labeling reagent (YMC CO, Kyoto, Japan), and then the amount of organic acid (lactic acid, acetic acid, propionic acid, butyric acid) was measured by HPLC. Furthermore, the pH of the filtered supernatant was also measured.
- YMC CO short-chain fatty acid analysis labeling reagent
- the amount of lactic acid and acetic acid in the feces of mice was significantly increased in the group fed with lactic acid bacteria powder compared to the control group (lactic acid: P ⁇ 0.01, acetic acid: P ⁇ 0.05).
- the pH of the group fed with lactic acid bacteria powder was significantly lower than that of the control group (P ⁇ 0.05).
- Example 5 Gene expression analysis of GRP43 The mouse obtained in (1) to (4) of Example 1 was euthanized by cervical dislocation 60 days after ingestion of lactic acid bacteria powder. Peyer's patch cells were collected and RNA was extracted using the illustra RNAspin Mini RNA Icolation kit (GE Healthcare, Piscataway, NJ, USA). 0.15 ⁇ g of the RNA was measured and reverse-transcribed using an oligo primer (Invitrogen, Carlsbad, Calif.) And Super Script III reverse transcriptase (Invitrogen).
- cDNA was purified from the reverse transcribed sample using a PCR Purification kit (Qiagen, Cambrige, MA, USA), and gene expression analysis of GRP43 in the intestinal tract was performed by a real-time PCR method using SYBR Green (Life Technologies).
- the endogenous control gene used in this example is ⁇ -actin as in Example 4. Further, the following primers were used for gene analysis.
- Statistical analysis was performed using Excel Statistics (SSRI, Tokyo, Japan). The data was subjected to statistical analysis by multiple comparison test (Dunnett's test).
- Example 6 When the lactic acid bacterium of the present invention or a processed product thereof was given to animals such as cattle (dairy cows, beef cattle), pigs, chickens (broilers, layers), etc., who had diarrhea, the diarrhea stopped and liquid feces solidified And so on. In addition, the amount of pharmaceutical feed required for normal breeding was reduced, and in the case of dairy cows and chickens, the occurrence of immune system diseases (eg, mastitis, colibacillosis, etc.) was suppressed. In broilers, the mortality rate was reduced. That is, the immunostimulatory action of the lactic acid bacteria of the present invention was confirmed.
- animals such as cattle (dairy cows, beef cattle), pigs, chickens (broilers, layers), etc.
- Example 7 When the lactic acid bacterium of the present invention or a processed product thereof was given to a healthy adult human with hay fever, the symptoms of hay fever were suppressed. That is, the antiallergic action of the lactic acid bacteria of the present invention is confirmed.
- Example 8 When the lactic acid bacteria of the present invention or a processed product thereof were given to rice, melon, tomato, or Chinese cabbage, the rice, melon, tomato fruit portion, and Chinese cabbage leaf were enlarged, and vitality was imparted to these plants. That is, the plant growth control action of the lactic acid bacteria of the present invention was confirmed.
- Example 9 When the lactic acid bacteria of the present invention or a processed product thereof were mixed and given to the animal feed of cattle (beef cattle), pigs, chickens (broilers), the feed requirement rate was reduced, compared to the case where feed containing these was not given, Changes such as firmness were observed. That is, the growth promoting effect of the lactic acid bacteria of the present invention on animals was confirmed.
- Example 10 When the lactic acid bacteria of the present invention or a processed product thereof were mixed with a fertilizer for paddy rice, the autoimmune action of the plant was exhibited and the whole was less likely to fall over when fertilizer not containing these was given. From this, it is thought that the lactic acid bacteria of this invention have a plant lodging prevention effect.
- Example 11 When the lactic acid bacteria of this invention or its processed material was mixed with the fertilizer of rice, the taste of rice increased compared with the case where the fertilizer which does not contain these was given. From this, it is considered that the lactic acid bacteria of the present invention have a plant taste-adjusting action.
- Example 12 According to a known method, the base sequence of Enterococcus dulans bacteria (E. durans HS-08 strain) of the present invention was determined and was as follows (SEQ ID NO: 13).
- the lactic acid bacteria of the present invention when administered to animals or mixed with feed, (1) promote the intestinal action of animals (livestock) and prevent infectious diseases and the like. Therefore, the amount of antibiotics used is reduced and the mortality rate is reduced. Furthermore, (2) the preference of animals (livestock) is increased, the growth promotion rate is improved, and the number of shipping days can be shortened. Moreover, (3) Since the odor due to the stool is reduced, the stress due to the barn environment is reduced and the meat quality is improved, which is useful.
- the lactic acid bacteria of the present invention are applied to a plant or mixed with a fertilizer, for example, (1) since vitality is imparted to the plant, the effect that the growth of the plant can be adjusted. Have. Moreover, (2) Since the autoimmunity action of a plant is exhibited, it has an effect of preventing plant lodging. Furthermore, (3) it has the effect of increasing the umami of the plant, so it is useful.
- the present invention provides a novel strain of Lactobacillus enterococcus durans (E. durans HS-08) or a treated product thereof and a method for growing the strain, and thus has industrial applicability.
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Abstract
Description
[1]エンテロコッカス・デュランス HS-08(受託番号NITE BP-02675)の菌体又はその処理物。
[2][1]に記載の菌体又はその処理物を含有する、IgA産生誘導用又は免疫賦活用組成物。
[3][1]に記載の菌体又はその処理物を含有する、抗アレルギー用組成物。
[4][1]に記載の菌体又はその処理物を含有する、有害菌及び/又は病原菌の増殖抑制用組成物。
[5][1]に記載の菌体又はその処理物を含有する、粘膜保護用組成物。
[6][1]に記載の菌体又はその処理物を含有する、短鎖脂肪酸産生の増強、有機酸産生の増強、及びGRP43の遺伝子発現上昇用 からなる群から選択される少なくとも1種の用途に用いられることを特徴とする組成物。
[7][1]に記載の菌体又はその処理物を含有する、植物の生長調節用、植物の倒伏防止用、及び植物の旨味調整用からなる群から選択される少なくとも1種の用途に用いられることを特徴とする組成物。
[8][1]に記載の菌体又はその処理物を含有する、動物の成長促進用及び/又は動物の自己免疫向上用の用途に用いられることを特徴とする組成物。
[9][1]に記載の菌体又はその処理物、又は[7]に記載の組成物を含有することを特徴とする植物用肥料。
[10][1]に記載の菌体又はその処理物、又は[8]に記載の組成物を含有することを特徴とする動物用飼料。
[11][1]に記載の菌体又はその処理物、又は[2]~[8]のいずれかに記載の組成物を含有することを特徴とする飲食品、化粧品又は医薬品。
[12][1]に記載の菌体又はその処理物、又は[2]~[8]のいずれかに記載の組成物を含有することを特徴とするコンクリート部材。
[13][1]に記載の菌体又はその処理物、並びにその他の乳酸菌、ビフィズス菌、若しくは酵母菌を含む有用菌、及びカビからなる群から選択される少なくとも1種又はそれらの処理物を含有する組成物。
[14]エンテロコッカス・デュランス HS-08(受託番号NITE BP-02675)の菌体を培養することを特徴とする、エンテロコッカス・デュランス HS-08の菌体の増殖方法。
本発明の乳酸菌は、免疫抗体の1種であるIgAの顕著な産生誘導作用及びそのことによる生体における免疫賦活作用を促す。
また、本発明の乳酸菌は、抗アレルギー作用、さらに驚くべきことに、有害菌及び/又は病原菌の増殖抑制作用、粘膜保護作用、短鎖脂肪酸産生の増強作用、有機酸産生の増強作用、GRP43の遺伝子発現上昇作用、植物の生長調節、倒伏防止、若しくは旨味調整作用又は動物の成長促進、若しくは自己免疫向上作用を示す。
(i)培養液(培地):MRS(de Man, Rogosa and Sharpe)培地、MRS寒天培地、LBS培地、変法LBS寒天培地、Rogosa培地、その他公知の乳酸菌用培地又は培養液等を含む。(ii)pHは、4~9程度、好ましくは、5~8程度、さらに好ましくは、5.5~7.5程度である。(iii)培養方法は、静置培養でもよいが、攪拌培養でもよい。攪拌培養の場合の攪拌回数は、1分間あたり、100~250回程度でよく、好ましくは、1分間あたり、120~180回程度、さらに好ましくは、1分間あたり、140~160回程度である。(iv)培養時間は、通常は1~72時間程度、好ましくは、3~48時間程度、さらに好ましくは、6~24時間程度である。(v)培養温度は、通常は20~40℃程度、好ましくは、25~39℃程度、さらに好ましくは、30~38℃程度である。(vi)その他、MRS培地、MRS寒天培地、LBS培地、変法LBS寒天培地、Rogosa培地、その他公知の乳酸菌用培地又は培養液は、他の添加剤を含んでもよい。
また、本発明の組成物が投与される動物は、特に限定されないが、例えば、ヒト、又はヒト以外の動物等であってもよい。ヒト以外の動物としては、特に限定されないが、例えば、牛、馬、豚、羊等の家畜類、イヌ、ネコ、ウサギ、ハムスター、リス、モルモット、ネズミ等のペット又は実験動物、いわし、まぐろ、サメ、マンボウ、エイ、あじ、いとう、ぶり、鮎、鯉、鯛、鯖、鮃、鰈、鰻、ウツボ、あんこう、ふぐ、カワハギ、熱帯魚等の魚類、海老、蟹、烏賊、帆立、牡蠣、栄螺等の魚介類、ニワトリ、ウズラ、シチメンチョウ、アヒル、ガチョウ、インコ、オウム等の鳥類、カブトムシ、クワガタ、モンシロチョウ、アゲハチョウ、カマキリ、バッタ、鈴虫、ダンゴムシ等の虫類、ヤモリ、カメレオン、イグアナ、トカゲ等のは虫類、イモリ、カエル、サンショウウオ等の両生類等が挙げられる。このうち、ヒト、牛、豚、ニワトリ、魚類、海老への投与が特に好ましい。
本発明は、本発明の乳酸菌若しくはその処理物、又はこれを含む組成物を含有する食品を提供する。食品には、健康食品、機能性表示食品、特定保健用食品、病者用食品が含まれる。食品の形態は特に限定されない。具体例には、これらに限定されないが、例えば、いわゆる栄養補助食品又はサプリメントとしての錠剤、顆粒剤、散剤、ドリンク剤等を挙げることができる。これ以外には、例えば、清涼飲料、栄養飲料、炭酸飲料、果実飲料、乳酸飲料、茶飲料等の飲料、そば、うどん、中華麺、即席麺等の麺類、チョコレート、スナック菓子、キャンディー、ガム、ビスケット、ゼリー、ジャム、クリーム等の菓子類、パン類、ソーセージ、かまぼこ、ハム等の水産加工食品、加工乳、発酵乳(ヨーグルトを含む)等の乳製品、マーガリン、マヨネーズ、ショートニング、ホイップクリーム、ドレッシング、サラダ油、てんぷら油等の油脂又は油脂加工食品、醤油、ソース等の調味料、カレー、丼、お粥、雑炊、シチュー等のレトルトパウチ食品、アイスクリーム、シャーベット等の冷菓などを挙げることができる。食品中のE. durans HS-08株の割合は、食品全体の質量に対し、好ましくは0.1~10%であり、より好ましくは1~10%である。
本発明は、本発明の乳酸菌若しくはその処理物、又はこれを含む組成物を含有する化粧品を提供する。化粧品には、いわゆる薬用化粧品(医薬部外品)が含まれる。化粧品としては、これらに限定されないが、例えば、洗浄剤、シャンプー、リンス、ヘアートニック、ヘアーローション、アフターシェーブローション、ボディーローション、化粧ローション、クレンジングクリーム、マッサージクリーム、エモリエントクリーム、エアゾール製品、消臭剤、芳香剤、脱臭剤又は入浴剤などを挙げることができる。本発明の化粧品は、本発明のレシチン以外に化粧品として一般に使用されている成分、例えば、界面活性剤、保湿剤、動植物由来油脂、微生物由来油脂、シリコーン類、高級アルコール、低級アルコール、動植物由来抽出エキス、微生物由来抽出エキス、紫外線吸収剤、消炎剤、金属封鎖剤、ビタミン類、酸化防止剤、増粘剤、防腐剤、殺菌剤、pH調整剤、着色剤、各種香料などを目的に応じて適宜配合することができる。これらについては、上記で例示したものを同様に用いることが出来る。化粧品中のE. durans HS-08株の割合は、化粧品全体の質量に対し、好ましくは0.1~10%であり、より好ましくは1~10%である。
本発明は、本発明の乳酸菌若しくはその処理物、又はこれを含む組成物を含有する医薬品を提供する。医薬品は、本発明の乳酸菌以外に、有効成分を含み、さらに薬学的に許容される担体、添加剤を必要に応じて適宜配合して製剤化することができる。医薬品は、これらに限定されないが、具体的には錠剤、被覆錠剤、丸剤、散剤、顆粒剤、カプセル剤、液剤、懸濁剤、乳剤、注射剤、輸液、坐剤、軟膏、スプレー剤、軟膏剤、クリーム剤、ゲル剤、パッチ剤等とすることができる。担体又は添加剤の配合割合については、医薬品分野において通常採用されている範囲に基づいて適宜設定すればよい。配合できる担体又は添加剤は、特に制限されないが、例えば、水、生理食塩水、その他の水性溶媒、水性又は油性基剤等の各種担体、賦形剤、結合剤、pH調整剤、崩壊剤、吸収促進剤、滑沢剤、着色剤、矯味剤、香料等の各種添加剤が挙げられる。これらの他、上記で例示したものを好ましく用いることも出来る。医薬品中のE. durans HS-08株の割合は、医薬品全体に対し、好ましくは0.1~10%であり、より好ましくは1~10%である。
本発明は、本発明の乳酸菌若しくはその処理物、又はこれを含む組成物を含有する動物用飼料を提供する。飼料としては、例えば、牛、馬、豚、羊等の家畜類用飼料、イヌ、ネコ、ウサギ、ハムスター、リス、モルモット、ネズミ等のペット又は実験動物類用飼料、いわし、まぐろ、サメ、マンボウ、エイ、あじ、いとう、ぶり、鮎、鯉、鯛、鯖、鮃、鰈、鰻、ウツボ、あんこう、ふぐ、カワハギ、熱帯魚等の魚類用飼料、海老、蟹、烏賊、帆立、牡蠣、栄螺等の魚介類用飼料、ニワトリ、ウズラ、シチメンチョウ、アヒル、ガチョウ、インコ、オウム等の鳥類用飼料、カブトムシ、クワガタ、モンシロチョウ、アゲハチョウ、カマキリ、バッタ、鈴虫、ダンゴムシ等の虫類用飼料、ヤモリ、カメレオン、イグアナ、トカゲ等のは虫類用飼料、イモリ、カエル、サンショウウオ等の両生類用飼料などが挙げられるが、これらに限定されない。本発明の飼料は、飼料中に本発明の乳酸菌を添加する以外、一般的な飼料の製造方法を用いて製造することができる。飼料中のE. durans HS-08株の割合は、飼料全体に対し、好ましくは0.1~10質量%であり、より好ましくは1~10質量%である。当該動物用飼料は、動物の成長促進又は自己免疫向上や、飼料を与えた動物(特に家畜又は家禽)の飼育期間短縮、又は早期出荷につながる。
本発明は、本発明の乳酸菌若しくはその処理物、又はこれを含む組成物を含有する植物用肥料を提供する。ここで、肥料としては、例えば、野菜、花、花木又は果物等が挙げられるが、これらに限定されない。また、例えば、野菜又は果物としては、大根、ゴボウ、人参、かぶ、レンコン等の根菜類、じゃがいも、サツマイモ、長芋、タロイモ、里芋、コンニャクイモ、山芋等のいも類、枝豆、空豆、エンドウ豆、インゲン、レンズ豆、小豆、ごま等の豆類、キャベツ、レタス、白菜、ほうれん草、野沢菜、水菜、三つ葉、小松菜、春菊等の葉野菜、蜜柑、甘夏、文旦、椪柑、伊予柑、八朔、レモン、ライム、グレープフルーツ等の柑橘類、玉葱、長ネギ、白ネギ等の葱類、きゅうり、えのき、えりんぎ、きくらげ、しめじ、なめこ、松茸、椎茸、舞茸等のきのこ類、ブロッコリー、カリフラワー、菜花、アーティチョーク等の花菜類、キュウリ、スイカ、カボチャ、ズッキーニ、ヒョウタン、ヘチマ、トウガン、テッポウウリ、ユウガオ、ニガウリ、ゴーヤー、メロン等のウリ類、水稲、ライ麦、鳩麦、きび、アワ、ひえ、もろこし等の稲類、小麦、大麦等の麦類、わらび、ぜんまい等の山菜、その他、ソテツ、イチョウ、マツ、竹、桃、クリ、柿、イチゴ、リンゴ、なし、ぶどう、マスカット、トマト、なす、わさび、セロリ、たけのこ、ニンニクなどが挙げられる。また、花又は花木としては、アザミ、アヤメ、カスミソウ、カタバミ、タンポポ、エンドウ、ホウセンカ、カーネーション、カモミール、サクラソウ、ユリ、サルビア、ボタン、アブラナ、キク、キンポウゲ、ヒガンバナ、パンジー、スミレ、ヒマワリ、ヒヤシンス、チューリップ、スイセン、フヨウ、スイートピー、マリーゴ-ルド、スズラン、ナデシコ、ガ-ベラ、ゼラニウム、ヒャクニチソウ、キキョウ、コスモス、ダリア、アサガオ、ヒルガオ、クロッカス、ケイトウ、アジサイなどの花、バラ、ウメ、サクラ、モモ、ボケ、スモモ、アンズ等のバラ科、キンモクセイ、ジャスミン、ヒイラギ、ライラック、レンギョウ、オウバイ等のモクセイ科、ハナミズキ等のミズキ科、ジンチョウゲ科、フジ、ハギ等のマメ科、ツバキ、サザンカ等のツバキ科、サツキ、ツツジ、シャクナゲ、アセビ、ブルーベリー等のツツジ科、その他、アオイ科、カエデ科、ジンチョウゲ科、エゴノキ科、トチノキ科、ウルシ科、スイカズラ科などの花木が挙げられるが、これらに限定されない。本発明の肥料は、肥料中に本発明の乳酸菌を添加する以外、一般的な肥料の製造方法を用いて製造することができる。肥料中のE. durans HS-08株 の割合は、肥料全体に対し、好ましくは0.1~10質量%であり、より好ましくは1~10質量%である。また、当該植物用肥料は、植物の生長調節、植物の倒伏防止、又は植物の旨味調整等の作用を有し得る。当該植物用肥料は、例えば、植物の根、茎、枝、葉、果実、又は花の生長を促進し得る。また、当該植物用肥料は、例えば、植物の根、茎、枝、葉、果実、又は花の旨味を増強し得る。
本発明は、本発明の乳酸菌若しくはその処理物、又はこれを含む組成物を含有する工業製品(例えば、コンクリート部材等)を提供する。コンクリート部材以外の工業製品としては、これらに限定されないが、例えば、石油製品、樹脂製品、磁気製品、皮革製品、織物などが挙げられる。工業製品中のE. durans HS-08株の割合は、工業製品に対し、好ましくは0.1~10質量%であり、より好ましくは1~10質量%である。
当該工業製品は、動物の成長促進、動物の自己免疫向上、植物の生長調節、植物の倒伏防止、又は植物の旨味調整等の作用を有し得る本発明の乳酸菌若しくはその処理物、又はこれを含む組成物を含有するため、動物の飼育に用いられる工業製品(例えば、餌箱、柵等)植物の生育に用いられる工業製品(例えば、鉢植え、じょうろ、バケツ、ホース、取水口のコンクリート等)である場合、動物の成長促進、動物の自己免疫向上、植物の生長調節、植物の倒伏防止、又は植物の旨味調整等に役立つ。
本発明では、本発明の乳酸菌若しくはその処理物、又はこれを含む組成物を含有するキットを提供する。キットは、例えば、分割式ボトル又はホイルパケットのように、組成物を別途含めることができる容器からなることが好ましいが、組成物を単一の容器に含めることもできる。好ましいキットの例としては、錠剤、カプセル等のパッケージングに用いられるブリスターパックや、薬剤液を充填してなる注射筒及び容器などを挙げることができるが、これらに限定されない。なお、キットは、放射線、オートクレーブなど公知の滅菌処理を行っても良い。
本発明の乳酸菌は、好ましくは、免疫賦活作用を有する。免疫賦活作用に関して、例えば、本発明の乳酸菌又は乳酸菌含有組成物の投与群が、非投与群と比較して、糞便中又は血液中IgA濃度が有意差又は有意傾向を示して高い場合、免疫賦活作用を有すると判断しうる。また、例えば、本発明の組成物が飼料に混ぜられて動物に投与された場合、免疫性疾患(例えば、乳房炎、大腸菌症、下痢等)の症状を緩和することが出来る。このことによって、抗生物質の使用量を低減することもできる。
また、粘膜組織の免疫賦活作用に関し、酢酸はレチノイン酸を誘導することで、IgAを誘導することが知られており、また、パイエル板の樹状細胞が分泌するIL-6や、RALDH2、APRIL、BAFF等もIgA産生を誘導することが知られている。そのため、例えば、これらが、本発明の乳酸菌又は乳酸菌含有組成物の投与群が、非投与群と比較して、酢酸濃度やRALDH2、IL-6、APRIL、BAFFの遺伝子発現量が有意差又は有意傾向を示して高い場合、粘膜組織の免疫を賦活し、感染症を予防する効果を有すると判断しうる。ここで、好ましくは、粘膜組織は、例えば、消化器官、呼吸器官、泌尿器等であり、さらに好ましくは、腸管の粘膜組織の免疫系が賦活されることが好ましい。
本発明の乳酸菌は、好ましくは、粘膜保護作用を有する。腸内細菌の産生する乳酸や酢酸等の有機酸は、ヒトを含む動物の様々な生体反応に寄与するが、そのような有機酸は、粘膜組織から吸収され、粘膜上皮細胞のエネルギー源となり、上皮細胞を修復して、粘膜を保護する作用を有することも知られている。そのため、例えば、本発明の乳酸菌又は乳酸菌含有組成物の投与群が、非投与群と比較して、粘膜組織中の乳酸や酢酸量が有意差又は有意傾向を示して高い場合、粘膜保護効果を有すると判断しうる。
本発明の乳酸菌は、好ましくは、抗アレルギー作用を有する。例えば、本発明の乳酸菌含有組成物の投与群が、非投与群と比較して、血液中又は糞中のIgA濃度が有意差又は有意傾向を示して高い場合、抗アレルギー作用を有すると判断しうる。また、例えば、本発明の組成物がヒトに投与された場合、花粉症等の症状を緩和することが出来る。
本発明の乳酸菌は、好ましくは、短鎖脂肪酸又は有機酸の産生増強作用を有する。IgAの産生には、乳酸菌等の腸内細菌の産生する短鎖脂肪酸又は有機酸が寄与することが知られているので、例えば、本発明の乳酸菌又は乳酸菌含有組成物の投与群が、非投与群と比較して、短鎖脂肪酸又は有機酸の濃度が有意差又は有意傾向を示して高い場合、短鎖脂肪酸又は有機酸の産生増強作用を有すると判断しうる。
本発明の乳酸菌が産生増強する短鎖脂肪酸又は有機酸としては、乳酸、酢酸などが挙げられるが、これらに限定されない。短鎖脂肪酸は、IgAによる免疫賦活作用や抗アレルギー作用をもたらすことが知られており、人や動物の生体に様々な好影響を与えると考えられる。
本発明の乳酸菌は、好ましくは、酢酸受容体GRP43の遺伝子発現上昇作用を有する。GRP43は短鎖脂肪酸、特に酢酸に対する親和性が高い受容体であり、腸内の短鎖脂肪酸量の増加に伴い、GRP43も増加することが知られている。つまり、GRP43の遺伝子発現量が増加すれば、腸管内の短鎖脂肪酸(酢酸等)が増加すると考えられる。
また、GPR43の遺伝子発現上昇は、インスリンシグナルの調節を介した糖尿病予防効果、恒常性維持に関与する制御性T細胞の誘導による大腸炎抑制効果など、様々な有用な作用を有することが知られている(Nature Communications volume 4, Article number: 1829 (2013))。そのため、例えば、本発明の乳酸菌又は乳酸菌含有組成物の投与群が、非投与群と比較して、GPR43遺伝子発現量が有意差又は有意傾向を示して高い場合、糖尿病予防効果や大腸炎抑制効果を有すると判断しうる。
本発明の乳酸菌は、好ましくは、有害菌及び/又は病原菌の増殖抑制作用を有する。ヒトやマウス等の動物に対する有害菌や病原菌は、酸性環境で増殖が抑制されることが知られているので、乳酸や酢酸などの有機酸が増加すれば、腸内環境が酸性化されて、pHが低下し、腸内の有害菌や病原菌の増殖が抑制されると考えられる。例えば、本発明の乳酸菌又は乳酸菌含有組成物の投与群が、非投与群と比較して、腸内のpHが有意差又は有意傾向を示して低い場合、有害菌及び/又は病原菌増殖抑制作用を有すると判断しうる。
なお、本発明で増殖を抑制出来る有害菌や病原菌としては、ウェルシュ菌、大腸菌等が挙げられるが、これらに限定されない。
本発明の乳酸菌は、好ましくは、植物の生長調節作用を有する。例えば、本発明の乳酸菌又は乳酸菌含有組成物の投与群が、非投与群と比較して、根、葉、枝、果実又は花の大きさにおいて生長著しい場合、例えば、これらの大きさが、好ましくは、10~100%程度、さらに好ましくは、30~50%程度生長した場合に、生長調節作用を有すると判断しうる。このことによって、植物生長のための肥料の使用量を低減することもできる。
本発明の乳酸菌は、好ましくは、動物の成長促進作用を有する。例えば、本発明の乳酸菌又は乳酸菌含有組成物の投与群が、非投与群の体重に対して、好ましくは、10~200質量%程度、さらに好ましくは、30~100質量%程度体重が増加することや、非投与群の体長に対して、好ましくは、10~200%程度、さらに好ましくは、30~100%程度体長が増加することがあるが、これらに限定されない。このことによって、飼料の使用量を低減することもできる。本開示において動物とは、例えば、上記したものが挙げられるが、これらに限定されない。
(1)供試マウスは、BALB/cAとし、入荷時の週齢は6週齢、性別はオスである。尚、入荷直後のマウスは、環境の変化によるストレスが原因となり、試験結果に影響を与える可能性があるため、精製飼料(AIN-76A、Research Diets, New Brunswick, NJ)のみを摂取させる順化期間を17日設け、その後は、以下で述べる飼料を投与する期間及び観察期間とした(約60日間)。
また、乳酸菌粉末を摂取させたグループ(0.38%グループ)は、血液中のIgA濃度がコントロールグループよりも高かった。血液中のIgA濃度の向上は、全身の免疫力の向上といえるため、E.durans HS-08株は全身の免疫系を賦活し得ることが確認された。
(1)順化期間経過後、乳酸菌粉末を摂取させた後から0日目、7日目、14日目、21日目、28日目、35日目、42日目、49日目、56日目及び60日目に体重を測定した。
また、コントロールグループ、乳酸菌粉末を摂取させたグループの両区分とも、IgEは検出されなかった。このことから、E.durans HS-08株はマウスに対して、アレルギー性を有しない、すなわち、アレルゲンではないと考えられる。
さらに、薬剤耐性試験やリムルス試験等も行ったが、いずれも本発明の乳酸菌の安全性が確認できた。上記の結果をまとめると、下記の表6の通りである。
実施例1の(1)~(4)で得たマウスを、乳酸菌粉末を摂取させてから60日目に、頚椎脱臼で安楽死させ、マウスのパイエル板細胞を採取し、RNAをillustra RNAspin Mini RNA Icolationキット (GE Healthcare, Piscataway, NJ, USA) を用いて抽出した。そのRNAを0.15μg測り取り、オリゴプライマー(Invitrogen, Carlsbad, CA)、Super ScriptIII 逆転写酵素 (Invitrogen)を用いて逆転写した。その後、PCR Purificationキット (Qiagen, Cambrige, MA, USA)を用いて逆転写したサンプルからcDNAを精製し、SYBR Green (Life Technologies)によるリアルタイムPCR法により、腸内における免疫誘導因子の遺伝子発現解析を行った。
実施例1の(1)~(4)で得たマウスを、乳酸菌粉末を摂取させてから60日目のマウスから1日分の糞を採取した後、その糞を凍結乾燥し、超純水で懸濁した。その後、4℃で1時間置いてから遠心し、その上清を濾過した。濾過した上清を短鎖脂肪酸分析用ラベル化試薬 (YMC CO, Kyoto, Japan) で処理した後、HPLCで有機酸量(乳酸、酢酸、プロピオン酸、酪酸)を測定した。さらに、濾過した上清のpHも測定した。
また、図9に示す通り、マウスの糞中において、乳酸菌粉末を摂取させたグループのpHは、コントロールグループと比較して有意に低下していた(P <0.05)。
実施例1の(1)~(4)で得たマウスを、乳酸菌粉末を摂取させてから60日目に、マウスを頚椎脱臼で安楽死させ、マウスのパイエル板細胞を採取し、RNAをillustra RNAspin Mini RNA Icolationキット (GE Healthcare, Piscataway, NJ, USA) を用いて抽出した。そのRNAを0.15μg測り取り、オリゴプライマー (Invitrogen, Carlsbad, CA)、Super Script III 逆転写酵素 (Invitrogen) を用いて逆転写した。その後、PCR Purificationキット(Qiagen, Cambrige, MA, USA)を用いて逆転写したサンプルからcDNAを精製し、SYBR Green (Life Technologies) によるリアルタイムPCR法で腸管内におけるGRP43の遺伝子発現解析を行った。本実施例で用いた内在性コントロール遺伝子は実施例4と同様、β-アクチンである。また、遺伝子解析用のプライマーは下記のものを用いた。
本願発明の乳酸菌又はその処理物を、下痢をしている牛(乳牛、肉牛)、豚、鶏(ブロイラー、レイヤー)等の動物に与えたところ、下痢が止まり、液状であった糞便が固形化等した。また、通常飼育の過程で必要な医薬品の飼料添加量が抑えられ、乳牛や鶏の場合に、免疫系疾患(例えば、乳房炎、大腸菌症等)の発生が抑えられた。また、ブロイラーにおいては、死亡率の低減がみられた。すなわち、本発明の乳酸菌の免疫賦活作用が確認された。
本願発明の乳酸菌又はその処理物を花粉症の健康な成人のヒトに与えたところ、花粉症の症状が抑えられた。すなわち、本発明の乳酸菌の抗アレルギー作用が確認される。
本願発明の乳酸菌又はその処理物を米、メロン、トマト、又は白菜に与えたところ、米、メロン、トマトの果実部分、及び白菜の葉が大きくなり、これらの植物に活力が付与された。すなわち、本願発明の乳酸菌の植物の生長調節作用が確認された。
本願発明の乳酸菌又はその処理物を牛(肉牛)、豚、鶏(ブロイラー)の動物の飼料に混ぜて与えたところ、これらを含まない飼料を与えた場合に対し、飼料要求率が減少し、体型がしっかりとする等の変化が認められた。すなわち、本願発明の乳酸菌の動物の成長促進作用が確認された。
本願発明の乳酸菌又はその処理物を水稲の肥料に混ぜたところ、これらを含まない肥料を与えた場合に対し、植物の自己免疫作用が発揮され、全体が倒れにくくなった。このことから、本願発明の乳酸菌は、植物の倒伏防止作用を有するものと考えられる。
本願発明の乳酸菌又はその処理物を米の肥料に混ぜたところ、これらを含まない肥料を与えた場合に対し、米の旨味が増した。このことから、本願発明の乳酸菌は、植物の旨味調整作用を有するものと考えられる。
また、本発明の乳酸菌が、例えば、植物に散布された場合や肥料に混ぜられた場合には、(1)植物に活力が付与されるため、植物の生長を調節することが出来るという効果を有する。また、(2)植物の自己免疫作用が発揮されるため、植物の倒伏防止効果を有する。さらに、(3)植物の旨味が増す効果を有するため、有用である。
Claims (14)
- エンテロコッカス・デュランス HS-08(受託番号NITE BP-02675)の菌体又はその処理物。
- 請求項1に記載の菌体又はその処理物を含有する、IgA産生誘導用又は免疫賦活用組成物。
- 請求項1に記載の菌体又はその処理物を含有する、抗アレルギー用組成物。
- 請求項1に記載の菌体又はその処理物を含有する、有害菌及び/又は病原菌の増殖抑制用組成物。
- 請求項1に記載の菌体又はその処理物を含有する、粘膜保護用組成物。
- 請求項1に記載の菌体又はその処理物を含有する、短鎖脂肪酸産生の増強、有機酸産生の増強、及びGRP43の遺伝子発現上昇用からなる群から選択される少なくとも1種の用途に用いられることを特徴とする組成物。
- 請求項1に記載の菌体又はその処理物を含有する、植物の生長調節用、植物の倒伏防止用、及び植物の旨味調整用からなる群から選択される少なくとも1種の用途に用いられることを特徴とする組成物。
- 請求項1に記載の菌体又はその処理物を含有する、動物の成長促進用及び/又は動物の自己免疫向上用の用途に用いられることを特徴とする組成物。
- 請求項1に記載の菌体又はその処理物、又は請求項7に記載の組成物を含有することを特徴とする植物用肥料。
- 請求項1に記載の菌体又はその処理物、又は請求項8に記載の組成物を含有することを特徴とする動物用飼料。
- 請求項1に記載の菌体又はその処理物、又は請求項2~8のいずれかに記載の組成物を含有することを特徴とする飲食品、化粧品又は医薬品。
- 請求項1に記載の菌体又はその処理物、又は請求項2~8のいずれかに記載の組成物を含有することを特徴とするコンクリート部材。
- 請求項1に記載の菌体又はその処理物、並びにその他の乳酸菌、ビフィズス菌、若しくは酵母菌を含む有用菌、及びカビからなる群から選択される少なくとも1種又はそれらの処理物を含有する組成物。
- エンテロコッカス・デュランス HS-08(受託番号NITE BP-02675)の菌体を培養することを特徴とする、エンテロコッカス・デュランス HS-08の菌体の増殖方法。
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| US12514755B2 (en) | 2020-03-05 | 2026-01-06 | Smith & Nephew Plc | Sensor integrated dressings and systems |
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| JP2023128590A (ja) * | 2022-03-03 | 2023-09-14 | 株式会社アルソア慧央グループ | 腸管免疫賦活剤、IgA産生促進剤及び遺伝子発現促進剤 |
| JP7717383B2 (ja) | 2022-03-03 | 2025-08-04 | 株式会社アルソア慧央グループ | 腸管免疫賦活剤、IgA産生促進剤及び遺伝子発現促進剤 |
| CN117586050A (zh) * | 2024-01-19 | 2024-02-23 | 山东嘉禾海洋生物科技有限公司 | 一种鱼蛋白发酵液及其制备方法和应用 |
| CN117586050B (zh) * | 2024-01-19 | 2024-04-02 | 山东嘉禾海洋生物科技有限公司 | 一种鱼蛋白发酵液及其制备方法和应用 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN112088211A (zh) | 2020-12-15 |
| JPWO2019230183A1 (ja) | 2021-06-17 |
| JP7012840B2 (ja) | 2022-01-28 |
| US20210205378A1 (en) | 2021-07-08 |
| KR102500894B1 (ko) | 2023-02-20 |
| KR20210005671A (ko) | 2021-01-14 |
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