WO2024090413A1 - 乳酸菌、該乳酸菌由来の自然免疫活性化剤及び該乳酸菌を含有する食品 - Google Patents

乳酸菌、該乳酸菌由来の自然免疫活性化剤及び該乳酸菌を含有する食品 Download PDF

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WO2024090413A1
WO2024090413A1 PCT/JP2023/038277 JP2023038277W WO2024090413A1 WO 2024090413 A1 WO2024090413 A1 WO 2024090413A1 JP 2023038277 W JP2023038277 W JP 2023038277W WO 2024090413 A1 WO2024090413 A1 WO 2024090413A1
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lactic acid
acid bacterium
acid bacteria
product
natural
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French (fr)
Japanese (ja)
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和久 関水
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Antenna Co Ltd
Genome Pharmaceuticals Institute Co Ltd
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Antenna Co Ltd
Genome Pharmaceuticals Institute Co Ltd
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A61K35/741—Probiotics
    • A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/04—Immunostimulants
    • 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; 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
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology

Definitions

  • the present invention relates to lactic acid bacteria, natural immune activators derived from said lactic acid bacteria, and foods containing said lactic acid bacteria.
  • Lactic acid bacteria have long been used in fermented foods and in the production of foods, medicines, probiotics, etc. Lactic acid bacteria are characterized by being gram-positive, catalase-negative, not forming endospores, and not being motile.
  • lactic acid bacteria are most commonly used as probiotics. Establishing an efficient method for isolating highly functional lactic acid bacteria and culturing them in foods will be useful for developing foods that use functional lactic acid bacteria.
  • Lactic acid bacteria are found in a variety of conventionally known foods, and there have been reports of isolating lactic acid bacteria from foods and identifying the species. Examples of such reports are as follows:
  • Non-patent literature 1 Leuconostoc mesenteroides (isolation source: dairy products)
  • Non-patent literature 2 Pediococcus ethanolidurans (isolation source: apple wine)
  • Non-patent document 3 Lactobacillus versmoldensis (isolated from sausage)
  • Non-patent literature 4 Companilactobacillus versmoldensis (isolation source: sausage)
  • Non-patent literature 5 Streptococcus salivarius (isolation source: cheese)
  • Natural immunity is a defense mechanism that quickly eliminates foreign substances (bacteria, viruses, cancer cells) from the body without relying on antibodies, and is shared by all living organisms, from insects to mammals.
  • innate immunity is the first line of defense against the host, triggering subsequent immune responses, including antibody production.
  • various stimuli stimulate the secretion of cytokines by immune cells such as macrophages, which eliminate invading pathogens and transmit information to other immune cells.
  • invertebrates such as insects do not have acquired immunity and rely solely on innate immunity to eliminate pathogens.
  • innate immune mechanisms of insects and mammals have many similarities.
  • insect blood cells called hemocytes phagocytose invading foreign substances in the same way as mammalian macrophages.
  • Toll-like receptor involved in the innate immune response in mammals is known to be highly homologous to the Toll receptor involved in the innate immune response in Drosophila.
  • the present inventors have developed an evaluation method that can easily measure innate immune activation responses in silkworms, which only have innate immune mechanisms (Patent Document 1, etc., Non-Patent Documents 6 and 7, etc.).
  • This evaluation method makes use of the fact that silkworms contract their muscles as a result of activation of innate immunity, and evaluates the innate immune activation ability based on the degree of muscle contraction.
  • the present inventors have shown that this evaluation method can be used to evaluate (screen) innate immune activators that have innate immune activation effects in vertebrates such as humans (Patent Document 1, etc.).
  • Patent Document 2 Non-Patent Document 8, Non-Patent Document 9, etc.
  • the present invention has been made in view of the above-mentioned background art, and aims to provide a lactic acid bacterium that has a high ability to activate natural immunity, and further to provide a natural immunity activator that contains the lactic acid bacterium, killed cells of the lactic acid bacterium, or a processed product of the lactic acid bacterium as an active ingredient, and a food product that contains the lactic acid bacterium or the natural immunity activator.
  • the present invention provides a lactic acid bacterium belonging to the genus Leuconostoc, which has a deposit number of NITE BP-03717 at the National Patent Microorganisms Depositary (NPMD) of the National Institute of Technology and Evaluation (NITE).
  • NPMD National Patent Microorganisms Depositary
  • NITE National Institute of Technology and Evaluation
  • the present invention also provides an innate immune activator comprising the lactic acid bacterium, killed lactic acid bacterium, or a processed product of the lactic acid bacterium as an active ingredient
  • the present invention provides a natural immune activator, characterized in that the processed product of the lactic acid bacteria is at least one processed product selected from the group consisting of fermented products, cultured products, concentrates, pastes, dried products, liquefied products, diluted products, crushed products, sterilized products, and extracts from cultures of lactic acid bacteria.
  • the present invention also provides a food product containing the lactic acid bacteria.
  • the present invention also provides a food product containing the natural immune activator.
  • the present invention also provides a method for producing food, which includes a fermentation step using the lactic acid bacteria.
  • the present invention can provide lactic acid bacteria with high innate immune activation ability.
  • the present invention can also provide an innate immune activator that contains the lactic acid bacteria, killed cells of the lactic acid bacteria, or a processed product of the lactic acid bacteria as an active ingredient, and a food product that contains the lactic acid bacteria or the innate immune activator.
  • the strain of lactic acid bacteria of the present invention has been reported to have been isolated from food, i.e., it has been consumed in the past. For this reason, it can be said that the safety of the lactic acid bacteria and innate immune activators of the present invention is ensured to a certain extent, and the hurdle for adding them to food is low.
  • the lactic acid bacteria of the present invention are lactic acid bacteria belonging to the genus Leuconostoc, and are assigned the accession number NITE BP-03717 at the National Patent Microorganisms Depositary (NPMD) of the National Institute of Technology and Evaluation (NITE).
  • NITE National Patent Microorganisms Depositary
  • the lactic acid bacteria may be referred to as "lactic acid bacteria #1-1,”"lactic acid bacteria #1-1 strain,”"#1-1,” etc. Lactic acid bacteria #1-1 will be described in detail below.
  • Lactic acid bacteria #1-1 was first isolated from rice bran (bran bed).
  • Lactic Acid Bacteria #1-1 The morphology of Lactic Acid Bacteria #1-1 is as follows:
  • Lactic Acid Bacteria #1-1 The physiological and chemical taxonomic properties of Lactic Acid Bacteria #1-1 are as follows:
  • ⁇ Glycerol - ⁇ Erythritol: - ⁇ D-Arabinose: - ⁇ L-Arabinose: + D-Ribose: + ⁇ D-Xylose: + ⁇ L-Xylose: - ⁇ D-Adonitol: - ⁇ -Methyl-D-xylopyranoside: - ⁇ D-Galactose: + ⁇ D-Glucose: + ⁇ D-Fructose: + ⁇ D-Mannose: - ⁇ L-Sorbose: - ⁇ L-Rhamnose: - ⁇ Dulcitol: - ⁇ Inositol: - ⁇ D-Mannitol: - ⁇ D-Sorbitol: - ⁇ -Methyl-D-mannopyranoside: - ⁇ -Methyl-D-glucopyranoside: + ⁇ N-Acety
  • Lactic Acid Bacteria #1-1 regarding 16S rDNA which is used as an index for molecular biological phylogenetic classification, is as shown in SEQ ID NO: 1 of the attached sequence listing. That is, the base sequence of the 16S rDNA region was amplified by PCR from the genomic DNA of Lactic Acid Bacteria #1-1, and analyzed by a sequencer, resulting in the discovery of a base sequence corresponding to almost the entire length of the 16S rDNA.
  • SEQ ID NO:1 The base sequence of SEQ ID NO:1 is set forth below.
  • Lactic Acid Bacterium #1-1 When the above base sequence was subjected to a homology search using NCBI BLAST analysis, the base sequence of the 16S rDNA region of Lactic Acid Bacterium #1-1 showed 99% homology with the base sequence of Leuconostoc mesenteroides subsp. jonggajibkimchii strain DRC1506 (accession number: NR_157602.1), which belongs to the Leuconostoc genus, and therefore Lactic Acid Bacterium #1-1 belongs to Leuconostoc mesenteroides. However, even when only the 16S rDNA region was compared, there was no complete agreement, and therefore Lactic Acid Bacteria #1-1 of the present invention is a different lactic acid bacteria strain from the above strains.
  • Lactic Acid Bacteria #1-1 is a novel microorganism belonging to the genus Leuconostoc.
  • Lactic Acid Bacterium #1-1 is a novel isolated microbial strain.
  • Lactic acid bacteria #1-1 was deposited domestically at the National Institute of Technology and Evaluation (NPMD) of the National Institute of Technology and Evaluation (hereinafter referred to as "NITE"), Room 122, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, Japan, and was deposited under the accession number: NITE P-03717 (deposit date: August 12, 2022).
  • NPMD National Institute of Technology and Evaluation
  • NITE National Institute of Technology and Evaluation
  • Lactic acid bacteria #1-1 was then submitted to the Patent Microorganisms Depository (NPMD) of the National Institute of Technology and Evaluation (NITE), Room 122, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, Japan, for an original deposit application, and an application was made to transfer the domestic deposit (original deposit date: August 12, 2022) to a deposit under the Budapest Treaty (transfer date (international deposit date): June 19, 2023). Viability was proven, and the application to transfer to a deposit under the Budapest Treaty (international deposit) was accepted. As a result, the deposit number: NITE BP-03717 was assigned.
  • the properties of the strain are easily mutated, so the lactic acid bacteria #1-1 of the present invention may not remain within the range of physiological properties shown above. Furthermore, such "mutations" include both natural and artificial mutations.
  • Lactic Acid Bacteria #1-1 may be cultured according to a general culture method used for microorganisms of the genus Leuconostoc. The culture is preferably carried out under anaerobic conditions.
  • Carbon sources in the medium include organic carbon compounds such as D-ribose, D-galactose, D-glucose, D-fructose, D-mannose, D-mannitol, N-acetylglucosamine, amygdalin, arbutin, esculin, salicin, D-cellobiose, D-maltose, sucrose, D-trehalose, gentiobiose, molasses, starch syrup, and fats and oils
  • nitrogen sources include organic and inorganic nitrogen compounds such as meat extract, casein, peptone, yeast extract, dry yeast, germ, soybean flour, urea, amino acids, and ammonium salts.
  • inorganic salts such as sodium salts, potassium salts, calcium salts, magnesium salts, phosphates, iron salts, copper salts, zinc salts, cobalt salts, etc. may be added as necessary.
  • growth promoters such as biotin, vitamin B1, cystine, methyl oleate, lard oil, etc.
  • antifoaming agents such as silicone oil, surfactants, etc. may be added.
  • the prepared medium for example, MRS medium, GAM medium, etc. are preferably used.
  • the culture conditions may be similar to those generally used for the culture of Leuconostoc microorganisms, as described above.
  • static culture is preferable.
  • static culture using a glass bottle with a lid may be used.
  • the culture temperature is preferably kept between 25° C. and 37° C., and more preferably between 30° C. and 37° C.
  • the culture pH is preferably around 7.
  • the culture period is a factor that varies depending on the medium composition used, the culture temperature, etc., but in the case of Lactic Acid Bacterium #1-1, a sufficient amount of the target product can be obtained by culture for preferably 12 to 72 hours, and more preferably 24 to 48 hours. It is also preferable to pick up colonies obtained by culture and perform single colony formation again on a medium.
  • Lactic acid bacteria #1-1 of the present invention has the ability to activate natural immunity both as the lactic acid bacteria themselves and as naturally or artificially mutated lactic acid bacteria thereof. That is, the present invention relates to a lactic acid bacterium belonging to the genus Leuconostoc having the accession number NITE BP-03717, or a naturally or artificially mutated lactic acid bacterium thereof, which has the ability to activate innate immunity.
  • the "natural immunity activation ability" is measured by the method described in the Examples below.
  • the "processed product of lactic acid bacteria” includes at least one processed product selected from the group consisting of fermented products, cultured products, concentrates, pastes, dried products, liquefied products, diluted products, crushed products, sterilized products, and extracts from cultured products of lactic acid bacteria.
  • the "dried products” include spray-dried products, freeze-dried products, vacuum-dried products, drum-dried products, etc.
  • the present invention also relates to "a natural immune activator containing as an active ingredient the lactic acid bacteria of the present invention (Lactic acid bacteria #1-1 or naturally or artificially mutated lactic acid bacteria thereof), killed cells of the lactic acid bacteria, or a processed product of the lactic acid bacteria, characterized in that the processed product of the lactic acid bacteria is at least one processed product selected from the group consisting of fermented products, cultured products, concentrates, pastes, dried products, liquefied products, diluted products, crushed products, sterilized products, and extracts from cultures of lactic acid bacteria.”
  • a natural immune activator containing as an active ingredient the lactic acid bacteria of the present invention (Lactic acid bacteria #1-1 or naturally or artificially mutated lactic acid bacteria thereof), killed cells of the lactic acid bacteria, or a processed product of the lactic acid bacteria, characterized in that the processed product of the lactic acid bacteria is at least one processed product selected from the group consisting of fermented products, cultured products, concentrates, pastes, dried products,
  • the innate immunity activator of the present invention can contain the lactic acid bacteria of the present invention, killed cells of the lactic acid bacteria, or a processed product of the lactic acid bacteria in various states, such as a suspension, lactic acid bacteria bodies, culture supernatant, or medium components.
  • the lactic acid bacteria that are the active ingredient of the innate immunity activator of the present invention may be in the form of live bacteria, wet bacteria, dried bacteria, etc.
  • the active ingredient of the innate immunity activator of the present invention may be killed bacteria obtained by sterilizing the lactic acid bacteria, i.e., by subjecting the lactic acid bacteria to heat sterilization, radiation sterilization, crushing, etc.
  • the content of the lactic acid bacteria, killed lactic acid bacteria, and processed product of the lactic acid bacteria, which are the active ingredients in the innate immune activator of the present invention, relative to the entire innate immune activator is not particularly limited and can be selected appropriately depending on the purpose, but when the entire innate immune activator is taken as 100 parts by mass, the total amount of "lactic acid bacteria, killed lactic acid bacteria, and processed product of the lactic acid bacteria" is preferably 0.001 parts by mass or more, more preferably 0.01 parts by mass or more, and particularly preferably 0.1 parts by mass or more. It is also preferably 100 parts by mass or less, more preferably 99 parts by mass or less, and particularly preferably 95 parts by mass or less.
  • the active ingredients may be used alone or in combination of two or more kinds.
  • the content ratio of each active ingredient in the natural immune activator is not particularly limited and can be appropriately selected according to the purpose.
  • the innate immune activator of the present invention contains the lactic acid bacteria, killed lactic acid bacteria, or a processed product of the lactic acid bacteria as active ingredients, and can contain "other ingredients" in addition to these active ingredients.
  • the "other components" in the innate immune activator are not particularly limited and can be appropriately selected according to the purpose within a range that does not impair the effects of the present invention, and examples of such components include pharma- ceutically acceptable carriers.
  • the dosage form of the innate immune activator of the present invention is not particularly limited and can be appropriately selected depending on the desired administration method, for example, as described below.
  • Specific examples include oral solid preparations (tablets, coated tablets, granules, powders, hard capsules, soft capsules, etc.), oral liquid preparations (oral liquid preparations, syrups, elixirs, etc.), injections (solvents, suspensions, etc.), ointments, patches, gels, creams, external powders, sprays, and inhalation powders.
  • Oral solid preparations can be produced by adding, for example, excipients to the active ingredients, and, if necessary, additives such as binders, disintegrants, lubricants, colorants, and flavorings and odorants, in a conventional manner.
  • excipients include lactose, sucrose, sodium chloride, glucose, starch, calcium carbonate, kaolin, microcrystalline cellulose, and silicic acid.
  • binders include water, ethanol, propanol, simple syrup, glucose liquid, starch liquid, gelatin liquid, carboxymethyl cellulose, hydroxypropyl cellulose, hydroxypropyl starch, methyl cellulose, ethyl cellulose, shellac, calcium phosphate, and polyvinylpyrrolidone.
  • Disintegrants include, for example, dry starch, sodium alginate, agar powder, sodium hydrogen carbonate, calcium carbonate, sodium lauryl sulfate, monoglyceride stearate, lactose and the like.
  • Lubricants include, for example, purified talc, stearates, borax, polyethylene glycol, and the like.
  • colorants include titanium oxide and iron oxide.
  • flavoring and fragrance agents include sucrose, orange peel, citric acid, tartaric acid, and the like.
  • Oral liquid preparations can be produced by adding additives such as flavorings, buffers, stabilizers, edible (processed) oils, and animal and vegetable oils to the active ingredients in a conventional manner.
  • flavoring agents include sucrose, orange peel, citric acid, and tartaric acid.
  • buffering agent is sodium citrate.
  • Stabilizers include, for example, tragacanth, gum arabic, gelatin, and the like.
  • a pH adjusting agent for example, a buffering agent, a stabilizer, an isotonicity agent, a local anesthetic agent, etc.
  • injections for subcutaneous, intramuscular, intravenous, etc. can be produced by conventional methods.
  • pH adjusters and buffers include sodium citrate, sodium acetate, and sodium phosphate.
  • stabilizer include sodium pyrosulfite, EDTA, thioglycolic acid, and thiolactic acid.
  • isotonic agents include sodium chloride and glucose.
  • local anesthetics include procaine hydrochloride, lidocaine hydrochloride, and the like.
  • Ointments can be prepared, for example, by combining the active ingredients with known bases, stabilizers, humectants, preservatives, etc., and mixing them in a conventional manner.
  • bases include liquid paraffin, white petrolatum, white beeswax, octyldodecyl alcohol, paraffin, and the like.
  • preservatives include methyl parahydroxybenzoate, ethyl parahydroxybenzoate, and propyl parahydroxybenzoate.
  • Patches can be produced, for example, by applying the above-mentioned ointment in the form of a cream, gel, paste, etc., to a known support in a conventional manner.
  • the support examples include woven fabrics made of cotton, staple fiber, and chemical fibers, nonwoven fabrics, films such as soft polyvinyl chloride, polyethylene, polypropylene, and polyurethane, and foam sheets.
  • the innate immunity activator of the present invention can be suitably used for, for example, individuals, bacteria, etc. that require activation of the innate immune mechanism.
  • the composition can be administered to, for example, individuals who need to maintain their health or recover from fatigue; individuals who need to prevent or treat cancer or lifestyle-related diseases; individuals infected with bacteria, fungi, viruses, etc.; and the like.
  • animals to which the innate immune activator of the present invention may be administered include humans; laboratory animals such as mice and rats; monkeys; horses; livestock such as cows, pigs, goats, and chickens; and pets such as cats and dogs.
  • the method of administration of the natural immune activator is not particularly limited and can be appropriately selected depending on the dosage form described above, and examples of such methods include oral administration, intraperitoneal administration, injection into the blood, injection into the intestine, etc. Among these, oral administration is preferred because it is simple and exerts the above-mentioned effects.
  • the dosage of the natural immune activator is not particularly limited and can be appropriately selected depending on the age, body weight, and desired degree of effect of the individual to be administered, but for example, the daily dosage for an adult is preferably 1 mg or more, more preferably 10 mg or more, and particularly preferably 100 mg or more, as the amount of active ingredient. Also, it is preferably 30 g or less, more preferably 10 g or less, and particularly preferably 3 g or less.
  • the timing of administration is also not particularly limited and can be appropriately selected depending on the purpose. For example, the agent may be administered prophylactically or therapeutically.
  • the present invention also relates to a food product containing the above-mentioned lactic acid bacteria (Lactic Acid Bacteria #1-1) and a food product containing the above-mentioned innate immunity activator.
  • the present invention also relates to a method for producing a food product, the method comprising a step of fermenting the food product using the above-mentioned lactic acid bacteria (Lactic Acid Bacteria #1-1).
  • a food containing the above-mentioned lactic acid bacteria (Lactic Acid Bacteria #1-1)
  • a food containing the above-mentioned innate immune activator or "a food produced by a food production method having a fermentation step using the above-mentioned lactic acid bacteria (Lactic Acid Bacteria #1-1)”
  • the term “food” includes “drinks.”
  • Lactic Acid Bacteria #1-1 is a lactic acid bacterium belonging to Leuconostoc mesenteroides. It has been reported that Leuconostoc mesenteroides has been isolated from food (dairy products) as described in Non-Patent Document 1. In other words, Leuconostoc mesenteroides can be said to be a lactic acid bacterium that has been consumed in the past.
  • the content of lactic acid bacteria and natural immune activator in the food of the present invention is not particularly limited and can be selected appropriately depending on the purpose and the form (type) of the food, but when the entire food is taken as 100 parts by mass, the above total amount is preferably 0.001 parts by mass or more, more preferably 0.01 parts by mass or more, and particularly preferably 0.1 parts by mass or more. It is also preferably 100 parts by mass or less, more preferably 99 parts by mass or less, and particularly preferably 95 parts by mass or less.
  • any one of the above may be used alone, or two or more may be used in combination.
  • two or more are used in combination, there is no particular restriction on the content ratio of each substance in the above food, and it can be selected appropriately according to the purpose.
  • the food of the present invention has the ability to activate natural immunity.
  • the present invention is also a food for activating natural immunity.
  • the food of the present invention can further contain "other ingredients" in addition to the natural immunity activator of the present invention described above.
  • other ingredients are not particularly limited and can be selected appropriately according to the purpose within a range that does not impair the effects of the present invention, and examples include various food ingredients.
  • content of the “other ingredients” is not particularly limited and can be selected appropriately according to the purpose.
  • the food of the present invention may be a general food, or it may be a so-called health food (health functional food, food for specified health uses, food with nutrient functions, food with functional claims, etc.).
  • the type of general food is not particularly limited and can be selected appropriately depending on the purpose, and examples include confectioneries such as jelly, candy, chocolate, biscuits, gummies, etc.; beverages such as green tea, black tea, coffee, soft drinks, etc.; dairy products such as fermented milk, yogurt, ice cream, lacto ice cream, etc.; vegetable and fruit processed products such as vegetable drinks, fruit drinks, jams, etc.; liquid foods such as soups; grain processed products such as breads, noodles, etc.; various seasonings, etc. There are no particular limitations on the method for producing these foods, and they can be produced, for example, according to any of the usual methods for producing various foods.
  • the type of health food is not particularly limited and can be appropriately selected depending on the purpose.
  • examples include oral solid preparations such as tablets, granules, and capsules; oral liquid preparations such as oral liquid preparations and syrups; and the like.
  • the method for producing the above oral solid preparation and oral liquid preparation is not particularly limited and can be appropriately selected depending on the purpose.
  • the food of the present invention is particularly useful as a functional food, health food, etc., intended for activating the natural immune system and building resistance to infectious diseases.
  • the production method can be carried out by a method well known to those skilled in the art.
  • Those skilled in the art can produce the desired food by appropriately combining a step of mixing the (killed) cells or treated products of the lactic acid bacteria of the present invention with other ingredients, a molding step, a sterilization step, a fermentation step, a baking step, a drying step, a cooling step, a granulation step, a packaging step, etc.
  • 0.5 g of rice bran was collected with 10 ⁇ L of Aze and spread on MRS agar medium containing calcium carbonate (CaCO 3 -MRS agar medium).
  • MRS agar medium containing calcium carbonate CaCO 3 -MRS agar medium
  • 1 mL of physiological saline was added to 0.5 g of rice bran to suspend it, and after leaving it to stand for a while, the supernatant was diluted 10 -3 or 10 -5 with physiological saline and spread on CaCO 3 -MRS agar medium with 10 ⁇ L of Aze, and each plate was anaerobically cultured at 30°C for 3 to 6 days. Anaeropack Kenki was used for the culture.
  • colonies that had formed a clear zone around them were cultured again on CaCO 3 -MRS agar medium, and it was confirmed that they formed a clear zone. These were selected as candidate strains of lactic acid bacteria.
  • 16S rDNA was amplified from colonies of candidate lactic acid bacteria strains by colony PCR, and the sequence was subjected to a homology search with 16S rDNA sequences of bacteria registered in a database using BLAST to identify the bacterial species with the highest sequence homology.
  • the above test samples were diluted to various concentrations, and 50 ⁇ L of each was injected into silkworm muscle specimens to measure muscle contraction values (C values).
  • Injection of 50 ⁇ L of saline and 0.2 mL of air served as negative and positive controls, respectively.
  • Dose-response curves were created using medical statistics software Prizm.
  • Table 1 shows the bacterial species and the measurement results of the natural immune activation ability for each candidate strain collected from rice bran (bran bed).
  • Table 2 shows the bacterial species and the values of innate immune activation ability of known lactic acid bacteria strains that the inventors have previously measured using the same method.
  • the values of natural immune activation ability shown in Tables 1 and 2 are relative values when the activity per 1 mg of sample in Public Example 5 (Lactococcus lactis) (U/mg) is set to 1.
  • the lactic acid bacteria of the present invention have a high ability to activate natural immunity, and are therefore widely used in the production of general foods, health foods, and other foods.
  • SEQ ID NO:1 is the base sequence of almost the entire length of the 16S rDNA of an unknown strain belonging to the genus Leuconostoc.

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PCT/JP2023/038277 2022-10-25 2023-10-24 乳酸菌、該乳酸菌由来の自然免疫活性化剤及び該乳酸菌を含有する食品 Ceased WO2024090413A1 (ja)

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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2006028047A (ja) * 2004-07-14 2006-02-02 Kikkoman Corp インターロイキン１２産生促進剤およびその製造法
JP2016086711A (ja) * 2014-10-31 2016-05-23 キリン株式会社 微生物又はｄｎａの免疫賦活活性を予測する方法
WO2018034203A1 (ja) * 2016-08-16 2018-02-22 株式会社ゲノム創薬研究所 新規乳酸菌、新規乳酸菌を有効成分として含有する自然免疫活性化剤、及び新規乳酸菌を含有する飲食品

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2006028047A (ja) * 2004-07-14 2006-02-02 Kikkoman Corp インターロイキン１２産生促進剤およびその製造法
JP2016086711A (ja) * 2014-10-31 2016-05-23 キリン株式会社 微生物又はｄｎａの免疫賦活活性を予測する方法
WO2018034203A1 (ja) * 2016-08-16 2018-02-22 株式会社ゲノム創薬研究所 新規乳酸菌、新規乳酸菌を有効成分として含有する自然免疫活性化剤、及び新規乳酸菌を含有する飲食品

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
MASAKI ISHII: "Lactic acid bacteria of the <i>Leuconostoc</i> genus with high innate immunity-stimulating activity", DD&T DRUG DISCOVERIES & THERAPEUTICS, INTERNATIONAL ADVANCEMENT CENTER FOR MEDICINE & HEALTH RESEARCH CO., LTD., JAPAN, vol. 11, no. 1, 1 January 2017 (2017-01-01), Japan , pages 25 - 29, XP093161345, ISSN: 1881-7831, DOI: 10.5582/ddt.2016.01078 *

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