WO2023054625A1 - Composition contenant de l'acide thréonique et procédé pour la produire - Google Patents

Composition contenant de l'acide thréonique et procédé pour la produire Download PDF

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WO2023054625A1
WO2023054625A1 PCT/JP2022/036544 JP2022036544W WO2023054625A1 WO 2023054625 A1 WO2023054625 A1 WO 2023054625A1 JP 2022036544 W JP2022036544 W JP 2022036544W WO 2023054625 A1 WO2023054625 A1 WO 2023054625A1
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bacteria
threonic acid
acid
genus
bifidobacterium
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PCT/JP2022/036544
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English (en)
Japanese (ja)
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一弥 戸田
真 吉本
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森永乳業株式会社
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Priority to JP2023551868A priority Critical patent/JPWO2023054625A1/ja
Publication of WO2023054625A1 publication Critical patent/WO2023054625A1/fr

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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 COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
    • A23L2/52—Adding ingredients
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
    • A61K31/19—Carboxylic acids, e.g. valproic acid
    • A61K31/191—Carboxylic acids, e.g. valproic acid having two or more hydroxy groups, e.g. gluconic acid
    • 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
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A61K35/741—Probiotics
    • A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
    • A61K35/745—Bifidobacteria
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A61K35/741—Probiotics
    • A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
    • A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P21/00—Drugs for disorders of the muscular or neuromuscular system
    • A61P21/06—Anabolic agents
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/40—Preparation of oxygen-containing organic compounds containing a carboxyl group including Peroxycarboxylic acids

Definitions

  • the present invention relates to a threonic acid-containing composition and a method for producing the same.
  • Muscles are the foundation of daily activities such as standing, walking, and maintaining posture. It exists regardless of gender. In addition, people who like to exercise, athletes, and the like desire to increase their muscle mass in order to improve their performance. Furthermore, as people become more health-conscious, how to increase muscle mass is becoming a social issue in improving quality of life (QOL), improving frailty, and preventing locomotive syndrome. It has become a challenge. In addition, “frailty” is a weakened state between a healthy state and a state in which life functions are impaired or a state requiring nursing care due to increased vulnerability to stress due to a decrease in physiological reserve in old age. It is a word indicating a state (Non-Patent Document 1).
  • Non-Patent Document 2 Muscle is mainly composed of protein and free amino acids. Also, branched-chain amino acids are known to act as stimuli to promote protein synthesis. Therefore, conventionally, foods and drinks containing various proteins and amino acids have been used to increase muscle mass (Non-Patent Document 2). In recent years, it has also been reported that ingestion of Bifidobacterium bacteria can thicken myotubes and increase muscle mass, and is expected to be a new active ingredient for increasing muscle mass ( Patent document 1).
  • an object of the present invention is to provide useful means for increasing muscle mass.
  • threonic acid activates muscle protein synthesis-related genes and promotes muscle hypertrophy. They also found that many Bifidobacterium bacteria and lactic acid bacteria produce threonic acid, and that culturing these bacteria in the presence of ascorbic acid promotes threonic acid production. Based on these findings, the inventors have conceived that threonic acid and threonic acid-producing bacteria can be used as an active ingredient in a composition for increasing muscle mass, and have completed the present invention.
  • the first aspect of the present invention provides bacteria of the genus Bifidobacterium, bacteria of the genus Lactobacillus, bacteria of the genus Lacticaseibacillus, bacteria of the genus Lactiplantibacillus, rimosi One or two or more bacteria selected from bacteria of the genus Limosilactobacillus, bacteria of the genus Levilactobacillus, bacteria of the genus Ligilactobacillus, and bacteria of the genus Latilactobacillus, ascorbic acids
  • a method for producing a threonic acid-containing composition or a composition for building muscle comprising the steps of culturing in a medium containing threonic acid and recovering a threonic acid-containing fraction from the culture after culturing.
  • the bacterium is preferably Bifidobacterium breve, more preferably Bifidobacterium breve FERM BP-11175.
  • a second aspect of the present invention is a combination of threonic acid and bacteria of the genus Bifidobacterium, Lactobacillus, Lacticaseibacillus, Lactipplantibacillus, Remocilactobacillus, and Levilactobacillus. , Rezilactobacillus, and Lactylactobacillus, and one or more selected from cultures of said bacteria. is.
  • a third aspect of the present invention relates to bacteria belonging to the genus Bifidobacterium, bacteria belonging to the genus Lactobacillus, bacteria belonging to the genus Lacticaseibacillus, bacteria belonging to the genus Lactiplantibacillus, bacteria belonging to the genus Remosilactobacillus, bacteria belonging to the genus Levilactobacillus, and bacteria belonging to the genus Lysilactobacillus.
  • compositions according to the third aspect are preferably used for increasing muscle mass.
  • compositions according to the second and third aspects of the present invention are hereinafter also referred to as "compositions of the present invention".
  • the bacterium is preferably Bifidobacterium breve, more preferably Bifidobacterium breve FERM BP-11175.
  • the composition of the present invention is preferably food or drink.
  • the compositions of the invention are preferably pharmaceuticals.
  • the present invention it is possible to provide a composition that can be taken continuously, is highly safe, and can effectively increase muscle mass. Therefore, by increasing muscle mass, daily life and exercise can be improved, and frailty and locomotive syndrome can be improved, so it is possible to support a healthy life in a wide range of age groups.
  • the present invention is a method for producing a composition for muscle building, comprising the steps of culturing a threonic acid-producing bacterium in an ascorbic acid-containing medium and recovering a threonic acid-containing fraction from the culture after culturing.
  • the first aspect of the present invention provides bacteria belonging to the genus Bifidobacterium, bacteria belonging to the genus Lactobacillus, bacteria belonging to the genus Lacticaseibacillus, bacteria belonging to the genus Lactiplantibacillus, bacteria belonging to the genus Remocilactobacillus, bacteria belonging to the genus Levilactobacillus, bacterium A step of culturing one or more bacteria selected from bacteria belonging to the genus Lactobacillus and bacteria belonging to the genus Lactobacillus in an ascorbic acid-containing medium, and recovering a threonic acid-containing fraction from the culture after culturing.
  • threonic acid-containing composition is not limited as long as it contains threonic acid, but contains any of bacterial cells, culture supernatant, medium, and purified threonic acid. Well, it may be a mixture of these. Further, the "threonic acid-containing composition” as used herein may contain a threonic acid-containing fraction described later.
  • NPMD Patent Microorganism Depositary Center
  • NITE Kazusa Kamatari, Kisarazu City, Chiba Prefecture 292-0818
  • IPTD is an abbreviation for the National Institute of Advanced Industrial Science and Technology Patent Organism Depositary Center. In 2012, the National Institute of Advanced Industrial Science and Technology inherited the international deposit status from the National Institute of Advanced Industrial Science and Technology to the National Institute of Technology and Evaluation, and the patent microorganism deposit business has been integrated into NPMD.
  • DSMZ is an abbreviation for Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH. The address is Inhoffenstr. 7B 38124 Braunschweig Germany. Strains with accession numbers beginning with “DSM” herein have been deposited with the DSMZ.
  • the following bacteria can be used when producing a threonic acid-containing composition containing bacterial cells.
  • the following bacteria can also be used when producing a threonic acid-containing composition containing any of culture supernatant, medium, and purified threonic acid.
  • Bifidobacterium bacteria include Bifidobacterium longum subspecies longum, Bifidobacterium longum subspecies infantis, Bifidobacterium breve, Bifidobacterium animalis sub Species animalis, Bifidobacterium animalis subspecies lactis, Bifidobacterium pseudolongum, Bifidobacterium pseudocatenulatum, Bifidobacterium adrecentis, and Bifidobacterium bifidum etc.
  • Bifidobacterium longum subspecies longum for example, Bifidobacterium longum subspecies longum NITE BP-02621 (alias: BB536 or Bifidobacterium longum subsp. longum ATCC BAA-999, for example, JP-A-2012- 223134, etc.), Bifidobacterium longum subspecies longum ATCC 15707, Bifidobacterium longum subspecies longum JCM1217 and the like can be used.
  • Bifidobacterium longum subspecies infantis for example, Bifidobacterium longum subspecies infantis M-63 (accession number NITE BP-02623), Bifidobacterium longum subspecies Species Infantis ATCC 15697 can be used.
  • Bifidobacterium breve for example, Bifidobacterium breve MCC1274 (accession number FERM BP-11175,) Bifidobacterium breve M-16V (accession number NITE BP-02622, commercially available, For example, "Bifidobacterium breve M-16V" manufactured by Morinaga Milk Industry Co., Ltd. may be used.), Bifidobacterium breve ATCC 15700, etc. may be used.
  • Bifidobacterium animalis subspecies animalis for example, Bifidobacterium animalis subspecies animalis ATCC25527 can be used.
  • Bifidobacterium pseudolongum for example, Bifidobacterium pseudolongum Subspecies pseudolongum JCM1205T can be used.
  • Bifidobacterium pseudocatenulatum for example, Bifidobacterium pseudocatenulatum DSM20438 can be used.
  • Bifidobacterium animalis subspecies lactis for example, Bifidobacterium animalis subspecies lactis DSM10140 can be used.
  • Bifidobacterium adolescentis JCM1275 can be used as Bifidobacterium adolescentis.
  • Bifidobacterium bifidum includes, for example, Bifidobacterium bifidum MCC1092 (accession number NITE BP-02429), Bifidobacterium bifidum MCC1319 (accession number NITE BP-02431), Bifidobacterium bifidum MCC1868 (Accession number NITE BP-02432), Bifidobacterium bifidum MCC1870 (accession number NITE BP-02433), Bifidobacterium bifidum JCM1255, etc. can be used.
  • Lactobacillus acidophillus Lactobacillus acidophillus, Lactobacillus acidophillus, Lactobacillus delbrueckii subsp. (Lactobacillus delbrueckii subsp. bulgaricus), Lactobacillus crispatus, Lactobacillus gasseri, Lactobacillus johnsonii, Lactobacillus helveticus, Lactobacillus amylovorus amylovorus).
  • Lactobacillus acidophilus Lactobacillus acidophilus MCC1847 (NITE BP-01695) can be used.
  • Lactobacillus gasseri Lactobacillus gasseri MCC1846 (NITE BP-01669), Lactobacillus gasseri JCM1131, and the like can be used.
  • Lactobacillus helveticus Lactobacillus helveticus JCM1120, Lactobacillus helveticus MCC1848 (NITE BP-01671), Lactobacillus helveticus MCC1844 (NITE BP-02185) and the like can be used.
  • the new classification of the genus Lactobacillus in the production method of the present invention (classified in the genus Lactobacillus in the old classification) includes Lacticaseibacillus casei (old classification name is Lactobacillus casei), Lacticaseibacillus paracasei (formerly classified as Lactobacillus paracasei), Lacticaseibacillus rhamnosus (formerly classified as Lactobacillus rhamnosus), and the like.
  • Lactobacillus casei for example, Lactobacillus casei JCM1134, Lactobacillus casei ATCC393, etc. can be used.
  • Lactobacillus paraceasi JCM8130 Lactobacillus paraceasi MCC1849 (NITE BP-01633), Lactobacillus paraceasi MCC1375 (NITE BP-11313) and the like can be used as Lactobacillus paraceasi.
  • Lactobacillus rhamnosus Lactobacillus rhamnosus JCM1136, Lactobacillus rhamnosus MCC1855 (LCS742, commercially available, for example, "LCS-742" manufactured by Morinaga Milk Industry Co., Ltd. can be used), lacto Bacillus rhamnosus ATCC53103, Lactobacillus rhamnosus ATCC53103, etc. can be used.
  • Lactiplantibacillus plantarum (old classification name is Lactiplantibacillus plantarum). Lactobacillus plantarum), Lactiplantibacillus pentosus (formerly known as Lactiplantibacillus pentosus). Lactiprantivibacillus plantarum ATCC14917 can be used as Lactiprantivibacillus plantarum.
  • Limosilactobacillus fermentum (old classification name) is Lactobacillus fermentum), and Limosilactobacillus reuteri (formerly classified as Lactobacillus reuteri).
  • Remosilactobacillus fermentum Remosilactobacillus fermentum SBS-1 can be used.
  • Remocilactobacillus reuteri JCM1112 Remocilactobacillus reuteri DSM17938, Remocilactobacillus reuteri ATCC PTA 6475, Remocilactobacillus reuteri ATCC PTA 5289, etc. can be used. .
  • Ligilactobacillus salivarius (formerly classified as lacto Bacillus salivarius).
  • Lactobacillus curvatus (formerly classified as Lactobacillus curvatus).
  • any one or a combination of two or more of the above bacteria can be used.
  • Bifidobacterium breve Bifidobacterium longum subspecies longum, Bifidobacterium longum subspecies infantis, Bifidobacterium animalis subspecies animalis, Bifidobacterium pseudolongum, Bifidobacterium pseudocatenulatum, Bifidobacterium adrecentis, Bifidobacterium animalis subspecies lactis, Lactobacillus gasseri, Lactobacillus helveticus, Lacti It is one or two or more selected from Bacillus casei, Lacticaseibacillus paracasei, Lacticaseibacillus rhamnosus, and Remocilactobacillus reuteri.
  • Bifidobacterium breve Bifidobacterium longum subspecies longum, Bifidobacterium longum subspecies infantis, Lactobacillus gasseri, Lactobacillus helveticus, Lacticaseibacillus It is one or two or more selected from Paracasei and Lacticaceae Bacillus rhamnosus. Even more preferred is Bifidobacterium breve or Bifidobacterium longum subspecies longum. Most preferred is Bifidobacterium breve FERM BP-11175 or Bifidobacterium longum NITE BP-02621.
  • the bacterium specified by the exemplified bacterial name is not limited to the strain itself (hereinafter also referred to as "deposited strain” for convenience of explanation) that has been deposited or registered with a predetermined institution under the bacterial name, Strains substantially equivalent thereto (also referred to as “derivative strains” or “derived strains”) are also included. That is, it is not limited to the strain itself deposited with the depositary institution under the above accession number, but also includes substantially equivalent strains.
  • a "strain substantially equivalent to the deposited strain” means that it belongs to the same species as the deposited strain, and the nucleotide sequence of the 16S rRNA gene is different from the nucleotide sequence of the 16S rRNA gene of the deposited strain. , preferably 97% or more, more preferably 98% or more, even more preferably 99% or more, even more preferably 100% identity, and preferably the same mycological properties as the deposited strain. refers to the shares held.
  • a strain substantially equivalent to the deposited strain may be, for example, a derivative of the deposited strain as a parent strain. Derivative strains include strains bred from the deposited strain and strains that arise naturally from the deposited strain.
  • Breeding methods include modification by genetic engineering techniques and modification by mutation treatment. Mutagenesis treatments include X-ray irradiation, ultraviolet irradiation, and treatment with mutating agents such as N-methyl-N'-nitro-N-nitrosoguanidine, ethyl methanesulfonate, and methyl methanesulfonate. be done. Strains naturally occurring from the deposited strain include strains naturally occurring during use of the deposited strain. Such strains include mutants naturally occurring by culturing (eg, subculturing) the deposited strain. Derivative strains may be constructed with one modification, or may be constructed with two or more modifications.
  • ascorbic acids are added to the medium when culturing the bacteria.
  • “ascorbic acids” means one or more selected from ascorbic acid, ascorbic acid derivatives, and salts thereof. L-isomers of ascorbic acids can usually be used. L-ascorbic acid can be used as ascorbic acid, and erythorbic acid, which is a stereoisomer of L-ascorbic acid, is also included.
  • ascorbic acid derivatives include salts of inorganic acid esters of L-ascorbic acid such as L-ascorbic acid phosphate and L-ascorbic acid sulfate, and L-ascorbic acid derivatives such as L-ascorbic acid-2-glucoside. Preferred examples include saccharides and the like. Salts of ascorbic acid or derivatives thereof are not particularly limited, but examples include alkali metal salts such as sodium salts and potassium salts, alkaline earth metal salts such as calcium and magnesium salts, ammonium salts, triethanolamine salts, triethylamine salts, and the like.
  • ascorbic acids in the present invention include sodium L-ascorbate, potassium L-ascorbate, calcium L-ascorbate, magnesium L-ascorbate, sodium erythorbate, potassium erythorbate, calcium erythorbate, and magnesium erythorbate. , L-ascorbic acid-2-glucoside, and the like.
  • the amount of ascorbic acid added is not particularly limited, but is preferably 1 mg/L to 100 g/L, more preferably 10 mg/L to 10 g/L, and still more preferably 100 mg/L to 1 g/L, relative to the medium. be able to. Also, the timing of adding ascorbic acids is not particularly limited, but it is usually added from the start of culture.
  • the culture step in the production method of the present invention is not particularly limited as long as the bacteria can grow.
  • the culturing method for example, the method normally used for culturing the bacteria can be used as it is or after being modified as appropriate.
  • the culture temperature may be, for example, 25-50°C, preferably 35-42°C.
  • Culturing is preferably carried out under anaerobic conditions, for example, while passing anaerobic gas such as carbon dioxide.
  • Cultivation can also be performed under microaerobic conditions such as liquid stationary culture. Cultivation can be carried out, for example, until threonic acid accumulates within and/or is secreted outside the cells to a desired extent, or until the bacteria proliferate to a desired extent. For example, it may be 12-72 hours.
  • the medium used for culture is not particularly limited as long as it contains ascorbic acids and the bacteria can grow.
  • a medium commonly used for culturing the bacteria can be used as it is or after being appropriately modified. That is, as carbon sources, for example, sugars such as galactose, glucose, fructose, mannose, cellobiose, maltose, lactose, sucrose, trehalose, starch, starch hydrolysate, blackstrap molasses, etc. can be used depending on the assimilation. .
  • ammonia ammonium salts such as ammonium sulfate, ammonium chloride and ammonium nitrate, and nitrates
  • inorganic salts include sodium chloride, potassium chloride, potassium phosphate, magnesium sulfate, calcium chloride, calcium nitrate, manganese chloride, and ferrous sulfate.
  • Organic ingredients such as peptone, soybean flour, defatted soybean meal, meat extract and yeast extract may also be used.
  • reinforced Clostridial medium Reinforced Clostridial medium
  • MRS medium de Man, Rogosa, and Sharpe medium
  • mMRS medium modified MRS medium
  • TOSP medium TOS propionate medium
  • TOSP Mup medium TOS propionate mupirocin medium
  • the production method of the present invention includes a step of collecting a threonic acid-containing fraction from the culture after the culturing step.
  • threonic acid-containing fraction may be any of bacterial cells, culture supernatant, medium, or a mixture thereof, as long as it contains threonic acid.
  • threonic acid is an active ingredient that exerts a muscle-increasing effect.
  • ascorbic acids to the medium and culturing threonic acid-producing bacteria, the presence of threonic acid was confirmed in the culture supernatant and bacterial cell crushing supernatant. It was clarified that it was metabolized, accumulated inside the cells, and also secreted outside the cells.
  • a step of treating the collected threonic acid-containing fraction by centrifugation, crushing, washing, freeze drying, spray drying, or the like is performed. may contain.
  • a crushed composition, washed composition, freeze-dried composition, or spray-dried composition is obtained. A process may be included.
  • the recovered threonic acid-containing fraction when producing a threonic acid-containing composition containing a culture supernatant, is treated by centrifugation, supernatant recovery, concentration, freeze drying, spray drying, or the like. may include a step of In the production method of the present invention, when producing a threonic acid-containing composition containing a culture supernatant, it includes a step of obtaining a concentrated composition, a freeze-dried composition, or a spray-dried composition. good too.
  • a step of treating the recovered threonic acid-containing fraction by centrifugation, medium recovery, concentration, freeze drying, spray drying, or the like is included. may contain.
  • a step of obtaining a concentrated composition, a freeze-dried composition, or a spray-dried composition when producing a threonic acid-containing composition containing a medium, it may include a step of obtaining a concentrated composition, a freeze-dried composition, or a spray-dried composition. .
  • a step of purifying threonic acid contained in the threonic acid-containing fraction may be included.
  • the step of purifying threonic acid is not particularly limited as long as it can purify threonic acid, but it is possible to use a step of crushing cells, centrifugation, membrane separation, solvent extraction, or the like.
  • the production method of the present invention can be a method for producing threonic acid.
  • the production method of the present invention When used as a method for producing threonic acid, it may contain a collected threonic acid-containing fraction.
  • the threonic acid-containing fraction recovered in this manner usually contains threonic acid of preferably 0.01 ⁇ g/g or more, more preferably 0.1 ⁇ g/g or more, and still more preferably 1 ⁇ g/g of threonic acid per total dry weight. g or more, still more preferably 5 ⁇ g/g or more, even more preferably 10 ⁇ g/g or more.
  • threonic acid usually means L-form.
  • threonic acid may be contained in the form of threonate, and examples of such salts include, but are not limited to, sodium salts, potassium salts, calcium salts, and the like.
  • the identification and quantification of threonic acid can be appropriately carried out using known analysis methods. For example, as shown in Examples below, it can be performed by liquid chromatography mass spectrometry (LC/MS). For example, Prominence (manufactured by Shimadzu Corporation) is used for HPLC, and TSQ Quantum Discovery MAX (Thermo company) can be used.
  • the production method of the present invention may include a step of treating the recovered threonic acid-containing fraction after the step of recovering the threonic acid-containing fraction.
  • the method of treating the threonic acid-containing fraction is not particularly limited as long as it does not impair the muscle-increasing action, and includes dilution, concentration, heating, freeze-drying, spray-drying, crushing, fractionation, and the like. That is, in the production method of the present invention, the steps of treating the recovered threonic acid-containing fraction include a step of separating and recovering bacterial cells, a step of separating and recovering the culture supernatant, a step of separating and recovering the medium, and a step of separating and recovering the medium containing threonic acid.
  • the production method of the present invention may be a step of purifying threonic acid from the fraction.
  • the production method of the present invention can be a method for producing threonic acid.
  • the production method of the present invention may include a step of mixing the collected threonic acid-containing fraction with the food raw material.
  • the production method of the present invention may include a step of converting the recovered threonic acid-containing fraction into a form such as powder, granules, paste, emulsion, wetting agent, capsule, tablet, or the like.
  • the production method of the present invention may include a step of sterilizing the recovered threonic acid-containing fraction after the step of recovering the threonic acid-containing fraction.
  • the step of sterilizing the threonic acid-containing fraction is not particularly limited as long as it does not impair the muscle volume-increasing action, and includes heat treatment, crushing treatment, pressure treatment, treatment with chemicals, and the like.
  • a step of sterilizing the cells may be included, and the sterilization may be heat treatment, crushing treatment, or pressure treatment. That is, when the resulting composition for increasing muscle mass contains bacterial cells as the threonic acid-containing fraction, the bacterial cells may be viable cells or dead cells. It may be a mixture of bodies.
  • the process of treating the recovered threonic acid-containing fraction, the process of mixing it with food ingredients, the process of making it into a liquid or the like, and the process of sterilizing are optional steps, and the order of the steps can be changed as appropriate. It is possible.
  • composition of the second aspect of the present invention comprises threonic acid, bacteria of the genus Bifidobacterium, bacteria of the genus Lactobacillus, bacteria of the genus Lacticaseibacillus, bacteria of the genus Lactipplantibacillus, bacteria of the genus Remocilactobacillus, and Reviract. containing one or more bacteria selected from bacteria belonging to the genus Bacillus, bacteria belonging to the genus Redilactobacillus, and bacteria belonging to the genus Rachilactobacillus, and one or more bacteria selected from cultures of the above bacteria to increase muscle mass provided for use.
  • a composition according to a second alternative aspect of the present invention contains purified threonic acid, or a threonic acid-containing fraction, for use in muscle building.
  • composition of the present invention can be preferably produced by the production method of the present invention described above.
  • threonic acid is usually included in the bacteria and/or culture thereof. Therefore, from another aspect, the composition of the present invention, as a third aspect, includes bacteria belonging to the genus Bifidobacterium, bacteria belonging to the genus Lactobacillus, bacteria belonging to the genus Lacticaseibacillus, bacteria belonging to the genus Lactipplantibacillus, and bacteria belonging to the genus Remocilactobacillus.
  • compositions can preferably serve for muscle building applications.
  • the type of bacteria in the composition of the present invention conforms to the description of the production method of the present invention described above.
  • the bacterium and/or its culture to be contained in the composition of the present invention may refer to the threonic acid-containing fraction recovered in the production method.
  • the content of the bacterium and/or its culture in the composition of the present invention is not particularly limited, but is, for example, 1.0 ⁇ 10 7 cfu or more per 1 g of the composition in terms of the amount of the bacterium. preferably 1.0 ⁇ 10 8 cfu or more, and still more preferably 1.0 ⁇ 10 9 cfu or more.
  • cfu refers to a colony forming unit. In this specification, for example, the value obtained when cultured at 38° C. in a solid medium containing 10% by mass of reconstituted skim milk can be used. Also, when the cells are dead cells, the cfu can be replaced with cells.
  • the content of the bacterial cells is preferably 0.1 mg or more, more preferably 1 mg or more, and still more preferably 10 mg or more per 1 g of the composition in terms of the amount of solids in the culture. .
  • the content of threonic acid in the composition of the present invention is not particularly limited, it is preferably 0.01 ⁇ g or more, more preferably 0.1 ⁇ g or more, and still more preferably 1 ⁇ g or more per 1 g of the composition. Preferably, it may contain 5 ⁇ g or more, and still more preferably 10 ⁇ g or more. These may be in the range of content when they are usually distributed as an oral composition.
  • the form of the composition of the present invention is not particularly limited. It may be a spray dried composition, a crushed composition, a fractionated composition, or a sterilized composition.
  • the form of the composition of the present invention may be liquid, powder, granules, paste, emulsion, wetting agent, capsule, tablet and the like.
  • the sterilization method is not limited, and it may be a composition sterilized by heat, a composition sterilized with a drug, or a composition sterilized by crushing. .
  • the composition of the present invention has the effect of increasing muscle mass in animals, including humans.
  • “Muscle mass” includes increasing muscle volume and/or weight and inhibiting muscle volume and/or weight loss. Increase in muscle volume and/or weight and inhibition of decrease in muscle volume and/or weight may be due to muscle hypertrophy (thickening of myotubes or muscle fibers), It may be due to an increase in the number of fibers.
  • the action of the composition of the present invention to increase muscle mass is believed to be due to the action of threonic acid activating protein synthesis-related genes such as p70S6K and promoting muscle protein synthesis.
  • composition of the present invention When the composition of the present invention is to be orally ingested (administered), it is preferably in the form of a food or drink. That is, another aspect of the present invention is the combination of threonic acid and Bifidobacterium, Lactobacillus, Lacticaseibacillus, Lactipplantibacillus, Remocilactobacillus, A composition containing one or more bacteria selected from bacteria belonging to the genus Levilactobacillus, bacteria belonging to the genus Rydilactobacillus, and bacteria belonging to the genus Lachilactobacillus, and one or more bacteria selected from cultures of the bacteria It is the use of things.
  • Another aspect of the present invention is the use of Bifidobacterium, Lactobacillus, Lacticaseibacillus, Lactipplantibacillus, Remocilactobacillus, Levilactobacillus, in muscle building.
  • Another aspect of the present invention is to provide bacteria of the genus Bifidobacterium, bacteria of the genus Lactobacillus, bacteria of the genus Lacticaseibacillus, bacteria of the genus Lactiplantibacillus, bacteria of the genus Remosilactobacillus, bacteria of the genus Levilactobacillus, and genus Lyzilactobacillus.
  • bacteria of the genus Bifidobacterium including administering to a subject one or more selected from bacteria and bacteria of the genus Lachilactobacillus, and one or more selected from cultures of said bacteria, said bacteria and/or said A method for increasing muscle bulk wherein the culture contains 1 ⁇ g/g or more of threonic acid per total dry weight thereof.
  • This specification also discloses, as a fourth aspect, a muscle building composition containing threonic acid as an active ingredient.
  • threonic acid activates protein synthesis-related genes in skeletal muscle cells, promotes muscle hypertrophy, and exerts a muscle-increasing effect.
  • Such invention can also be translated into the use of threonic acid in muscle mass building.
  • This invention can also be translated into a method for increasing muscle mass, including administering threonic acid to a subject.
  • the food and drink composition of the present invention can increase muscle mass, it prevents and/or improves symptoms such as muscle atrophy and diseases caused by decreased muscle mass, and prevents decreased muscle mass due to lack of exercise and aging. Prevention and/or amelioration is expected.
  • Diseases accompanied by symptoms such as muscle atrophy include, for example, hypotonia, muscle atrophy, muscle dystrophy, muscle degeneration, inflammatory muscle disease, myasthenia and sarcopenia.
  • Diseases caused by decreased muscle mass include osteoporosis, bone fracture, diabetes, chronic obstructive pulmonary disease, chronic kidney disease, dementia, and the like.
  • the food and drink composition of the present invention is also useful for healthy people and people who play sports because they can maintain and improve their daily lives and improve exercise efficiency.
  • "amelioration" of symptoms or diseases includes curing of diseases, alleviation of symptoms, reduction in severity of diseases or symptoms, and delay of progression of diseases or symptoms.
  • "prevention" of a symptom or disease includes preventing the occurrence of the symptom or disease, delaying the occurrence, and reducing the risk of the occurrence.
  • the subject (administrator) to whom the food and drink compositions of the second to fourth aspects of the present invention are administered (administrator) and the subject (consumer) to be ingested are not particularly limited as long as they are animals, but are usually humans. Moreover, any of adults, children, infants, newborns (including low-weight infants), and the like may be used. Moreover, sex is not specifically limited.
  • administering to a subject may be synonymous with “making a subject ingest”.
  • the intake is usually voluntary (free intake), but may be forced (forced intake).
  • the intake (administration) timing of the food and drink composition of the present invention is not particularly limited, and can be appropriately selected according to the condition of the intake (administration) subject.
  • the intake (administration) amount of the food and drink composition of the present invention is appropriately selected depending on the age, sex, condition, other conditions, etc. of the intake (administration) subject.
  • the intake (administration) amount of the food and drink composition of the present invention is preferably in the range of 100 ⁇ g/day to 10 g/day, for example, 1 mg/day in terms of the solid content of the bacteria and/or culture thereof for adults.
  • a range of -1 g/day is more preferred, and 10 mg/day to 500 mg/day is even more preferred.
  • the range for adults is preferably 1 ng/day to 100 ⁇ g/day, more preferably 1 ng/day to 1 ⁇ g/day, and even more preferably 10 ng/day to 500 ng/day.
  • the food and drink composition of the present invention can be ingested (administered) once a day or divided into multiple times a day, regardless of the amount and period of ingestion (administration).
  • the intake (administration) period of the food and drink composition of the present invention is not particularly limited. In addition, there is no particular upper limit for the intake (administration) period, and continuous long-term intake (administration) is possible.
  • the form and properties are not particularly limited as long as they can be orally ingested (administered) without impairing the effects of the present invention. It can be produced by a normal method using raw materials that are usually used for food and drink.
  • Food and drink regardless of the form such as liquid, paste, gel-like solid, powder, etc., for example, nutritional supplements (supplements), tablet confectionery; liquid diet (nutrition for ingestion); bread, macaroni, spaghetti Flour products such as , noodles, cake mixes, fried chicken powder, bread crumbs; instant noodles, cup noodles, retort/prepared foods, cooked canned foods, microwave oven foods, instant soups/stews, instant miso soups/soups, canned soups, freeze-dried foods , and other instant foods; canned agricultural products, canned fruits, jams and marmalades, pickled vegetables, boiled beans, dried agricultural products, processed agricultural products such as cereals (processed grains); canned marine products, fish meat hams and sausages , fish paste products, marine delicacies, processed fish products such as tsukudani; processed livestock products such as canned livestock products, pastes, meat hams and sausages; processed milk, milk drinks, yogurts, lactic acid bacteria drinks, cheese, ice creams Milk and dairy products
  • confectionery such as caramels, candies, chewing gums, chocolates, cookies, biscuits, cakes, pies, snacks, crackers, Japanese confectionery, rice confectionery, bean confectionery, dessert confectionery, jelly, and other confectionery; carbonated drinks, natural fruit juices , fruit juice drinks, soft drinks containing fruit juice, fruit drinks, fruit drinks containing fruit, vegetable drinks, soy milk, soy milk drinks, coffee drinks, tea drinks, powdered drinks, concentrated drinks, sports drinks, nutritional drinks, alcoholic beverages, etc.
  • Pleasant beverages such as beverages, other commercially available foods such as baby food, furikake, ochazuke seaweed, etc.; powdered milk for infants; enteral nutrition food; ) and the like.
  • the feed can also be set as feed as one aspect
  • the feed include pet food, livestock feed, fish feed, and the like.
  • the form of the feed is not particularly limited, and in addition to threonic acid, the bacteria and/or culture thereof, grains such as corn, wheat, barley, rye, and milo; soybean meal, rapeseed meal, coconut oil meal, and linseed meal.
  • Rice brans such as wheat bran, wheat bran, rice bran, and defatted rice bran; Manufacturing lees such as corn gluten meal and corn jam meal; Animal feeds such as fish meal, skimmed milk powder, casein, yellow grease, and tallow.
  • Yeasts such as torula yeast and brewer's yeast; mineral feed such as tribasic calcium phosphate and calcium carbonate; oils and fats; simple amino acids;
  • composition of the present invention is in the form of a food or drink (including feed), it can be provided and sold as a food or drink labeled with uses related to increasing muscle mass.
  • Such "display” acts include all acts for informing consumers of the above-mentioned use. Regardless of the object, medium, etc. to be displayed, all of them fall under the act of "display” in the present invention.
  • the "indication” be expressed in such a way that the consumer can directly recognize the use.
  • the act of transferring, handing over, displaying for the purpose of transfer or delivery, importing products related to food and beverages or product packaging that describes the above-mentioned use, advertisements related to products, price lists or transaction documents Examples include the act of displaying or distributing information describing the use, or providing information containing such information using an electromagnetic method (such as the Internet) after describing the use.
  • the content of the display is a display approved by the government (for example, a display that is approved based on various systems established by the government and performed in a manner based on such approval).
  • a display that is approved based on various systems established by the government and performed in a manner based on such approval it is preferable to attach such display contents to packaging, containers, catalogs, pamphlets, POP and other advertising materials at sales sites, other documents, and the like.
  • labeling includes labeling as health food, functional food, enteral nutrition food, food for special dietary use, food with health claims, food for specified health use, food with nutrient function claims, food with function claims, quasi-drugs, etc. is also mentioned.
  • the labeling approved by the Consumer Affairs Agency for example, the labeling approved by the system related to food for specified health use, food with nutrient function claims, or food with function claims, or similar system.
  • labeling as a food for specified health use labeling as a food for specified health use with certain conditions, labeling to the effect that it affects the structure and function of the body, labeling to reduce the risk of disease, labeling for functionality based on scientific evidence. Labeling, etc.
  • Such indications include, for example, “Increase muscle”, “For those who want to build muscle”, “For prevention of frailty”, “For improvement of QOL”, “Support for increasing muscle strength of exercisers”, and the like. mentioned.
  • One or more bacteria selected from bacteria belonging to the genus Bacillus, bacteria belonging to the genus Levilactobacillus, bacteria belonging to the genus Regilactobacillus, and bacteria belonging to the genus Lactobacillus, and bacteria belonging to the genus Lactobacillus, and one or more selected from cultures of the bacteria is a composition containing Another aspect of the present invention is the use of threonic acid and Bifidobacterium, Lactobacillus, Lacticaseibacillus, Lactipplantibacillus, Remocilactobacillus in the manufacture of a muscle building composition.
  • One or two or more bacteria selected from the genus bacteria, Levilactobacillus bacteria, Regilactobacillus bacteria, and Rachilactobacillus bacteria, and one or two or more bacteria selected from the cultures of the bacteria is used.
  • Another aspect of the present invention is the combination of threonic acid with Bifidobacterium, Lactobacillus, Lacticaseibacillus, Lactipplantibacillus, Remocilactobacillus, used for muscle gain.
  • Bacteria one or more bacteria selected from the genus Levilactobacillus, bacteria of the genus Regilactobacillus, and bacteria of the genus Lachilactobacillus, and one or more bacteria selected from cultures of the bacteria It is a composition that Another aspect of the present invention is the use of threonic acid with bacteria of the genus Bifidobacterium, Lactobacillus, Lacticaseibacillus, Lactipplantibacillus, Remocilactobacillus, Levilactobacillus, muscle, including administering to a subject one or more bacteria selected from bacteria belonging to the genus Rezilactobacillus and bacteria belonging to the genus Lachilactobacillus, and one or more bacteria selected from the cultures of said bacteria is a method of increasing the amount of Another aspect of the present invention is Bifidobacterium, Lactobacillus, Lacticaseibacillus, Lactipplantibacillus, Remosilactobacillus, Reviract used for muscle
  • Another aspect of the present invention is the use of Bifidobacterium, Lactobacillus, Lacticaseibacillus, Lactipplantibacillus, Remocilactobacillus, Levi in the manufacture of a muscle building composition.
  • Another aspect of the present invention is to provide bacteria of the genus Bifidobacterium, bacteria of the genus Lactobacillus, bacteria of the genus Lacticaseibacillus, bacteria of the genus Lactiplantibacillus, bacteria of the genus Remosilactobacillus, bacteria of the genus Levilactobacillus, and genus Lyzilactobacillus.
  • bacteria of the genus Bifidobacterium including administering to a subject one or more selected from bacteria and bacteria of the genus Lachilactobacillus, and one or more selected from cultures of said bacteria, said bacteria and/or said A method for increasing muscle bulk wherein the culture contains 1 ⁇ g/g or more of threonic acid per total dry weight thereof.
  • This specification also discloses, as a fourth aspect, a muscle building composition containing threonic acid as an active ingredient.
  • threonic acid activates protein synthesis-related genes in skeletal muscle cells, promotes muscle hypertrophy, and exerts a muscle-increasing effect.
  • Such an invention can also be rephrased as threonic acid used for increasing muscle mass.
  • Such an invention can also be translated into the use of threonic acid in the manufacture of a muscle building composition.
  • This invention can also be translated into a method for increasing muscle mass, including administering threonic acid to a subject.
  • the pharmaceutical composition of the present invention can increase muscle mass, it prevents and/or improves symptoms such as muscle atrophy and diseases caused by decreased muscle mass, and prevents decreased muscle mass due to lack of exercise and aging. and/or are expected to make improvements.
  • Diseases accompanied by symptoms such as muscle atrophy include, for example, hypotonia, muscle atrophy, muscle dystrophy, muscle degeneration, inflammatory muscle disease, myasthenia and sarcopenia.
  • Diseases caused by decreased muscle mass include osteoporosis, bone fracture, diabetes, chronic obstructive pulmonary disease, chronic kidney disease, dementia, and the like.
  • lack of exercise and age-related loss of muscle mass can cause sarcopenia, frailty, locomotive syndrome, and the like, so it is also expected to prevent and/or improve these.
  • the subject (administrator) to whom the pharmaceutical compositions of the second to fourth aspects of the present invention are administered (administrator) and the subject (ingester) to be ingested are not particularly limited as long as they are animals, but are usually humans. Moreover, any of adults, children, infants, newborns (including low-weight infants), and the like may be used. Moreover, sex is not specifically limited.
  • the timing of ingestion (administration) of the pharmaceutical composition of the present invention is not particularly limited, and can be appropriately selected according to the condition of the subject of ingestion (administration).
  • the intake (administration) amount of the pharmaceutical composition of the present invention is appropriately selected depending on the age, sex, condition, other conditions, etc. of the intake (administration) target.
  • the ingestion (administration) amount of the pharmaceutical composition of the present invention is preferably in the range of 100 ⁇ g/day to 10 g/day, and 1 mg/day to 1 mg/day in terms of solid content of the bacteria and/or culture thereof. A range of 1 g/day is more preferred, and 10 mg/day to 500 mg/day is even more preferred.
  • the range for adults is preferably 1 ng/day to 100 ⁇ g/day, more preferably 1 ng/day to 1 ⁇ g/day, and even more preferably 10 ng/day to 500 ng/day.
  • the pharmaceutical composition of the present invention can be ingested (administered) once a day or divided into multiple times a day, regardless of the amount and duration of ingestion (administration).
  • the timing of ingesting (administering) the drug of the present invention is not particularly limited, such as before meals, after meals, between meals, and before bedtime.
  • the intake (administration) period of the pharmaceutical composition of the present invention is not particularly limited. In addition, there is no particular upper limit for the intake (administration) period, and continuous long-term intake (administration) is possible.
  • the route of intake (administration) of the drug may be oral or parenteral, but oral is preferred.
  • parenteral intake (administration) includes percutaneous, intravenous, rectal administration, inhalation, and the like.
  • the pharmaceutical form it can be appropriately formulated into a desired dosage form depending on the intake (administration) method.
  • solid formulations such as powders, granules, tablets and capsules; and liquid formulations such as solutions, syrups, suspensions and emulsions can be formulated.
  • parenteral intake (administration) it can be formulated into suppositories, ointments, injections, and the like.
  • ingredients such as excipients, pH adjusters, colorants, and corrigents that are commonly used for formulation can be used.
  • other medicinal ingredients in combination, such as other medicinal ingredients, ingredients known or discovered in the future that have muscle-enhancing effects.
  • formulation can be appropriately carried out by a known method depending on the dosage form.
  • a carrier that is usually used for formulation may be blended as appropriate to form the formulation.
  • Such carriers include excipients, binders, disintegrants, lubricants, stabilizers, flavoring agents and the like.
  • Excipients include, for example, sugar derivatives such as lactose, sucrose, glucose, mannitol, sorbitol; starch derivatives such as corn starch, potato starch, ⁇ -starch, dextrin, carboxymethyl starch; crystalline cellulose, hydroxypropyl cellulose, Hydroxypropylmethylcellulose, carboxymethylcellulose, cellulose derivatives such as carboxymethylcellulose calcium; gum arabic; dextran; pullulan; silicate derivatives such as light silicic anhydride, synthetic aluminum silicate, and magnesium aluminometasilicate; phosphate derivatives such as calcium phosphate; carbonate derivatives such as calcium; sulfate derivatives such as calcium sulfate;
  • binders examples include gelatin; polyvinylpyrrolidone; macrogol, etc., in addition to the aforementioned excipients.
  • disintegrants include, in addition to the excipients mentioned above, chemically modified starch or cellulose derivatives such as croscarmellose sodium, carboxymethyl starch sodium, and crosslinked polyvinylpyrrolidone.
  • Lubricants include, for example, talc; stearic acid; metal stearates such as calcium stearate and magnesium stearate; colloidal silica; waxes such as Veegum and Geiro; ; carboxylic acid sodium salts such as sodium benzoate; sulfates such as sodium sulfate; leucine; lauryl sulfates such as sodium lauryl sulfate and magnesium lauryl sulfate; silicic acid anhydride and silicic acid hydrate; be done.
  • stabilizers include paraoxybenzoic acid esters such as methylparaben and propylparaben; alcohols such as chlorobutanol, benzyl alcohol and phenylethyl alcohol; benzalkonium chloride; acetic anhydride; and sorbic acid.
  • Flavoring agents include, for example, sweeteners, acidulants, flavoring agents, and the like.
  • examples of carriers used include solvents such as water.
  • Rat myoblast cell line L6 cells obtained from the American Type Culture Collection (ATCC) were adjusted to 1.5 ⁇ 10 4 cells/cm 2 .
  • the cells were seeded on a 6-well plate and cultured in DMEM medium (containing 10% Fetal Bovine Serum and 1% Penicillin Streptomycin) under 5% CO 2 at 37° C. for 24 hours.
  • DMEM medium containing 10% Horse Serum and 1% Penicillin Streptomycin; hereinafter referred to as "differentiation medium” was replaced with fresh medium every 2 days and cultured for 7 days.
  • the 1 M threonic acid sample prepared in (1) above was added to a final concentration of 1 or 10 ⁇ M, cultured for 1 hour, and then a lysate was prepared using RIPA buffer.
  • Activation (phosphorylation) of the protein synthesis-related gene p70S6K was assessed by Western blotting. Specifically, the fluorescence intensity of phosphorylated p70S6K and total p70S6K bands was measured using ChemiDoc (registered trademark) MP Imaging System (Bio-Rad Laboratories), and the ratio of phosphorylated p70S6K to total p70S6K was calculated. The activity value of p70S6K was used. As a control, the same procedure as described above was performed except that the threonic acid sample was not added.
  • HE staining hematoxylin and eosin staining
  • Tables 1 and 2 show the activity values of the protein synthesis-related gene (p70S6K) in L6 to which threonic acid was added.
  • the activity (degree of phosphorylation) of p70S6K increased 3.37-fold in the group treated with 1 ⁇ M threonate and increased 3.03-fold in the group treated with 10 ⁇ M threonate compared to the control.
  • Table 3 shows the diameters (average values) of myotube cells obtained by the procedure in (3) above. It was confirmed that the diameter of myotube cells increased by 2.58 ⁇ m or more in the 1 ⁇ M threonic acid-treated group and by 1.85 ⁇ m or more in the 10 ⁇ M threonic acid-treated group, compared to the control, and muscle mass increased.
  • 1% (v/v) of each was seeded in the MRS liquid medium and anaerobically cultured at 37°C (preculture). After 16 hours of culture, 1% (v/ v) were inoculated individually and anaerobically cultured at 37°C (main culture). After 16 hours of culture, the culture supernatant was collected by centrifugation. Bacteria were cultured in the same manner as a control, except that ascorbic acid was not added.
  • Test Example 3 Investigation of muscle volume-increasing effect by probiotic culture supernatant Using probiotic culture supernatant cultured in the presence of ascorbic acid, promotion of activity of protein synthesis-related gene (p70S6K) in L6 was evaluated, and the muscle volume-increasing effect was examined.
  • Test Example 2 The three strains prepared in Test Example 2 (1) (Bifidobacterium breve FERM BP-11175, Bifidobacterium pseudolongum subspecies pseudolongum JCM1205T, Limo Sodium hydroxide was added to the culture supernatant of Silactobacillus reuteri JCM1112T) to adjust the pH to 7.0 ⁇ 0.05, followed by filter sterilization.
  • Results Table 5 shows the activity value of the protein synthesis-related gene (p70S6K) in L6 of each group.
  • p70S6K activity values were higher when cultured in the presence of ascorbic acid than when cultured in the absence of ascorbic acid.
  • Culture supernatant of Bifidobacterium breve FERM BP-11175, Bifidobacterium pseudolongum subspecies pseudolongum JCM1205T, and Remocilactobacillus reuteri JCM1112T cultured in the presence of ascorbic acid was found to have a stimulatory effect on muscle protein synthesis.
  • ⁇ Test Example 4 Investigation of intracellular threonic acid production by probiotics (1) Culture of probiotics and quantification of threonic acid In the same procedure as in Test Example 2, each of the probiotics shown in Table 6 was cultured. bottom. The culture was centrifuged to collect the cells, washed with ultrapure water, suspended at a concentration of 10 mg/mL (wet weight of bacteria/ultrapure water), and treated with FastPrep-24 5G Homogenizer (manufactured by MP Biomedicals)). The resulting crude lysate was centrifuged to collect the supernatant (hereinafter referred to as "supernatant of cell lysate"). After that, in the same procedure as in Test Example 2, the amount of threonic acid in the supernatant of cell disruption was measured.

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Abstract

La présente invention vise à procurer un moyen utile pour augmenter la masse musculaire. La solution selon l'invention pour augmenter la masse musculaire consiste en une composition contenant de l'acide thréonique, une ou plusieurs bactéries choisies parmi Bifidobacterium spp., Lactobacillus spp., Lacticaseibacillus spp., Lactiplantibacillus spp., Limosilactobacillus spp., Levilactobacillus spp., Ligilactobacillus spp., et Latilactobacillus spp., et une ou plusieurs cultures des bactéries susmentionnées. Une telle composition est produite, par exemple, par un procédé comprenant une étape de culture d'une ou plusieurs bactéries choisies parmi les bactéries susmentionnées dans un milieu contenant de l'acide ascorbique et une étape de récupération d'une fraction contenant de l'acide thréonique à partir de la culture après culture.
PCT/JP2022/036544 2021-09-29 2022-09-29 Composition contenant de l'acide thréonique et procédé pour la produire WO2023054625A1 (fr)

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Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2016166176A (ja) * 2009-11-12 2016-09-15 ネステク ソシエテ アノニム 腸内微生物叢バランス及び健康を促進するための栄養組成物
WO2019087280A1 (fr) * 2017-10-31 2019-05-09 森永乳業株式会社 Composition pour augmentation de la masse musculaire
JP2019518744A (ja) * 2016-05-27 2019-07-04 ネステク ソシエテ アノニム 運動障害を治療又は予防するための栄養組成物

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2016166176A (ja) * 2009-11-12 2016-09-15 ネステク ソシエテ アノニム 腸内微生物叢バランス及び健康を促進するための栄養組成物
JP2019518744A (ja) * 2016-05-27 2019-07-04 ネステク ソシエテ アノニム 運動障害を治療又は予防するための栄養組成物
WO2019087280A1 (fr) * 2017-10-31 2019-05-09 森永乳業株式会社 Composition pour augmentation de la masse musculaire

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Title
KAZUYA TODA, YAMAUCHI YUKI, TANAKA AZUSA, KUHARA TETSUYA, ODAMAKI TOSHITAKA, YOSHIMOTO SHIN, XIAO JIN-ZHONG: "Heat-Killed Bifidobacterium breve B-3 Enhances Muscle Functions: Possible Involvement of Increases in Muscle Mass and Mitochondrial Biogenesis", NUTRIENTS, vol. 12, no. 1, 15 January 2020 (2020-01-15), pages 1 - 13, XP055745828, DOI: 10.3390/nu12010219 *

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