WO2018016393A1 - Composition destinée à la conservation d'acide désoxyribonucléique et procédé de production de ladite composition, procédé de conservation d'acide désoxyribonucléique, produit de bactéries lactiques et procédé d'utilisation dudit produit de bactéries lactiques - Google Patents

Composition destinée à la conservation d'acide désoxyribonucléique et procédé de production de ladite composition, procédé de conservation d'acide désoxyribonucléique, produit de bactéries lactiques et procédé d'utilisation dudit produit de bactéries lactiques Download PDF

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WO2018016393A1
WO2018016393A1 PCT/JP2017/025335 JP2017025335W WO2018016393A1 WO 2018016393 A1 WO2018016393 A1 WO 2018016393A1 JP 2017025335 W JP2017025335 W JP 2017025335W WO 2018016393 A1 WO2018016393 A1 WO 2018016393A1
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preserving
composition
lactic acid
acid bacteria
deoxyribonucleic acid
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PCT/JP2017/025335
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English (en)
Japanese (ja)
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雅史 森藤
晶美 北出
深澤　朝幸
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株式会社 明治
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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
    • 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
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
    • 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
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/20—Milk; Whey; Colostrum
    • 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
    • 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
    • A61K35/747—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
    • A61P17/00—Drugs for dermatological disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/18—Antioxidants, e.g. antiradicals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P39/00—General protective or antinoxious agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P39/00—General protective or antinoxious agents
    • A61P39/06—Free radical scavengers or antioxidants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • 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
    • 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
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/04—Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds

Definitions

  • the present invention relates to a composition for storing deoxyribonucleic acid (hereinafter referred to as “DNA”) and a method for producing the same.
  • the present invention also relates to a method for storing DNA.
  • the present invention further relates to lactic acid bacteria products and methods of use thereof.
  • An object of the present invention is to provide a composition for preserving DNA that can secure a sufficient supply amount and can keep the production cost low.
  • the composition for preserving DNA comprises a lactic acid bacterium product containing a polysaccharide (hereinafter referred to as “polysaccharide-containing lactic acid bacterium product”) as an active ingredient. That is, the polysaccharide-containing lactic acid bacteria product is used as a DNA storage composition or as one component thereof.
  • the “composition” referred to herein includes animals such as pharmaceuticals, supplements, food additives and the like, foods and drinks (excluding animals and plants themselves), and food and drink compositions (including processed foods and drinks). Included are those that can be ingested (including humans).
  • the term “preservation” as used herein means that DNA damage is suppressed or DNA damage repair is promoted.
  • the DNA storage composition according to the first aspect of the present invention can suppress DNA damage or promote DNA damage repair. That is, this DNA preserving composition can preserve DNA satisfactorily.
  • save composition is produced by lactic acid bacteria as above-mentioned. For this reason, the active ingredient of this composition for DNA preservation
  • a polysaccharide-containing lactic acid bacteria product is produced by a combination of Lactobacillus delbrueckii subsp. Bulgaricus and Streptococcus thermophilus.
  • the Lactobacillus delbruxii subspecies bulgaricus is specifically preferably Lactobacillus delbruxii subspecies bulgaricus OLL1247 (accession number: NITE BP-01814)
  • the thermophilus is specifically preferably Streptococcus thermophilus OLS3078 (accession number: NITE BP-01697).
  • the method according to the second aspect of the present invention is a method of using a polysaccharide-containing lactic acid bacteria product as a DNA storage composition. That is, it is to provide a polysaccharide-containing lactic acid bacteria product for use as a DNA storage composition.
  • the method for preserving DNA according to the third aspect of the present invention is a method for ingesting the above-described DNA preserving composition.
  • the intake period is preferably 1 week or longer, more preferably 2 weeks or longer, further preferably 3 weeks or longer, and particularly preferably 4 weeks or longer.
  • a composition for preserving DNA is produced by supplying a milk raw material to a lactic acid bacterium that produces a polysaccharide. That is, here, the polysaccharide-containing lactic acid bacteria product is used to produce a composition for preserving DNA.
  • FIG. 5 is a bar graph showing the abundance of 8-oxo-2'-deoxyguanosine in DNA collected from each sample according to an experimental example. It is a bar graph which shows the relative quantity of the mRNA of catalase measured using the cDNA synthesize
  • XPA xerderma pigmentmentum completion group A
  • a composition for preserving DNA is an orally ingested composition for suppressing DNA damage or promoting DNA damage repair, which is a lactic acid bacteria product containing a polysaccharide.
  • polysaccharide-containing lactic acid bacteria product is an active ingredient.
  • the polysaccharide-containing lactic acid bacteria product includes a polysaccharide-containing lactic acid bacteria fermented product (hereinafter referred to as “polysaccharide-containing lactic acid bacteria fermented product”), polysaccharide.
  • Lactic acid bacteria cultures hereinafter referred to as “polysaccharide-containing lactic acid bacteria cultures”
  • lactic acid bacteria metabolites containing polysaccharides and the like.
  • polysaccharide referred to here is a sugar chain polymer composed of saccharides such as galactose, glucose, rhamnose, mannose, N-acetylglucosamine.
  • the polysaccharides may include acidic polysaccharides to which phosphate groups are bonded.
  • the molecular weight is usually in the range of 5000 to 500,000.
  • lactic acid bacterium as used herein is a general term for microorganisms that assimilate glucose and produce lactic acid with a yield of sugar of 50% or more. It is a gram-positive staphylococci or gonococci with no motility. It has characteristics such as lack of spore formation and negative catalase. Lactic acid bacteria have been eaten all over the world through fermented milk since ancient times, and can be said to be extremely safe microorganisms.
  • lactic acid bacteria have been genus Lactococcus, Lactobacillus, Leuconostoc, Pediococcus, Streptococcus, Wissella, Tetrageno
  • the genus is classified into 11 genera such as Tetragenococcus genus, Oenococcus genus, Enterococcus genus, Vagococcus genus and Carnobacterium genus. All of these lactic acid bacteria can be used in the embodiment of the present invention.
  • Lactobacillus delbrueckii subspices bulgaricus Lactobacillus delbrueckii subspices bulgaricus (Lactobacillus delbrueckii subsp.
  • Lactobacillus delbruecki subspecies bulgaricus is preferably Lactobacillus delbrucky subspecies bulgaricus OLL1247 (accession number: NITE BP-01814)
  • Streptococcus thermophilus is Streptococcus -Thermophilus (Streptococcus thermophilus) OLS3078 (accession number: NITE BP-01697) is preferable.
  • lactic acid bacteria fermented product means a culture itself obtained by fermentation with lactic acid bacteria.
  • the fermented lactic acid bacterium includes a fermented lactic acid bacterium and a processed product thereof, for example, a culture filtrate or culture supernatant obtained by sterilizing a culture (fermented lactic acid bacterium) by filtration, centrifugation, membrane separation, or the like.
  • Liquid, culture filtrate / culture supernatant, lactic acid bacteria fermented product, and the like concentrated by an evaporator or the like, pasted product, diluted product (dried, heated, reduced pressure, etc.) dried product.
  • the preparation of the treated product should be performed by combining one or more of the aforementioned treatment steps such as sterilization treatment such as filtration, centrifugation, membrane separation, precipitation, concentration, pasting, dilution, drying, etc. Can do.
  • sterilization treatment such as filtration, centrifugation, membrane separation, precipitation, concentration, pasting, dilution, drying, etc.
  • the culture medium include nonfat dry milk medium and MRS medium to which yeast extract is added.
  • the lactic acid bacteria product is a milk fermentation product, a milk culture, or a milk metabolite of the lactic acid bacteria.
  • the fermented milk product, the milk culture product, and the milk metabolite include fermented milk (yogurt) and polysaccharides.
  • Fermented milk (yogurt) is a fermented food prepared by mixing lactic acid bacteria and yeast with milk and fermenting it, and is widely eaten from the viewpoint of its deliciousness, beauty and health. Fermented milk (yogurt) can be preferably used as its supernatant.
  • This fermented milk may be added with a thickening agent such as pectin, guar gum, xanthan gum, carrageenan, processed starch, or a gelling agent, in addition to a culture solution such as skim milk powder or a whey degradation product.
  • a thickening agent such as pectin, guar gum, xanthan gum, carrageenan, processed starch, or a gelling agent
  • milk examples include animal milk such as cow milk and processed products thereof (for example, skim milk, whole milk powder, skim milk powder, spinach, casein, whey, fresh cream, compound cream, butter, buttermilk powder, cheese Etc.), vegetable milk such as soybean milk derived from soybean.
  • animal milk such as cow milk and processed products thereof (for example, skim milk, whole milk powder, skim milk powder, spinach, casein, whey, fresh cream, compound cream, butter, buttermilk powder, cheese Etc.)
  • vegetable milk such as soybean milk derived from soybean.
  • the milk may be sterilized or may not be sterilized.
  • the fermented milk raw material mix is a mixture containing raw milk and other components.
  • This fermented milk raw material mix includes raw materials commonly used in the production of fermented milk such as raw milk, water, and other optional components (eg, sugar, sugar, sweetener, sour agent, mineral, vitamin, flavor, etc.). It is obtained by warming to dissolve and mixing.
  • Raw milk may contain water, raw milk, pasteurized milk, skim milk, whole milk powder, skim milk powder, whole fat concentrated milk, skim concentrated milk, butter milk, butter, cream, cheese, and the like.
  • the raw milk may contain whey protein concentrate (WPC), whey protein isolate (WPI), ⁇ -lactalbumin ( ⁇ -La), ⁇ -lactoglobulin ( ⁇ -Lg) and the like. .
  • fermented milk undergoes processes such as raw material mix preparation process, raw material mix (heating) sterilization process, raw material mix cooling process, starter addition process, fermentation process, fermented milk cooling process, etc.
  • raw material mix preparation step raw materials are mixed (prepared).
  • raw material mix cooling step raw material mix cooling step
  • starter addition step raw material mix cooling step
  • fermentation step fermented milk cooling step
  • a commonly used medium can be used. That is, any medium can be used as long as it contains a nitrogen source, an inorganic substance and other nutrients in addition to the main carbon source.
  • a nitrogen source lactose, glucose, sucrose, fructose, starch hydrolysate, molasses, etc. can be used depending on the assimilation ability of the bacteria used.
  • organic nitrogen-containing substances such as casein hydrolyzate, whey protein hydrolyzate, ⁇ -lactalbumin, ⁇ -lactoglobulin, glycomacropeptide, soybean protein hydrolyzate and the like can be used.
  • meat extract, fish extract, yeast extract and the like can be used as the growth promoter.
  • the lactic acid bacteria are preferably cultured in an anaerobic state, but are usually preferably cultured in a microaerobic state used in liquid stationary culture or the like.
  • a culture method under anaerobic conditions a known method such as a method of culturing in a carbon gas gas phase can be adopted, but other methods may be adopted.
  • the culture temperature is preferably in the range of 30 ° C. to 47 ° C., more preferably in the range of 35 ° C. to 46 ° C., and still more preferably in the range of 37 ° C. to 45 ° C. .
  • the pH of the medium during the cultivation of lactic acid bacteria is preferably maintained within the range of 6 or more and 7 or less, but may be other pH ranges as long as the pH allows the bacteria to grow.
  • the culture time for lactic acid bacteria and the like is usually preferably in the range of 1 hour to 48 hours, more preferably in the range of 1.5 hours to 36 hours, and more preferably in the range of 2 hours to 24 hours. More preferably it is.
  • Fermented milk typically has a non-fat milk solid content of 8% by weight or more, and the number of lactic acid bacteria or yeast is in the range of 10 6 / ml to 10 11 / ml.
  • composition for preserving DNA is a preparation such as pharmaceuticals, supplements and food additives, food and drink (except animals and plants themselves), and food and drink. It may take the form of a product composition (including processed food and drink).
  • the preparation is prepared as an oral preparation according to a conventional method using additives that are acceptable for formulation.
  • This preparation may take the form of solid preparations such as tablets, powders, fine granules, granules, capsules, pills, sustained-release preparations, and liquid preparations such as solutions, suspensions and emulsions.
  • Additives that are acceptable for formulation include, for example, excipients, stabilizers, preservatives, wetting agents, emulsifiers, lubricants, sweeteners, colorants, fragrances, buffers, antioxidants, pH Examples thereof include regulators.
  • Specific examples of food additives include seasonings such as processed seasonings, flavor seasonings, and cooking mixes.
  • the food and drink and the food and drink composition are processed for human and animal food and drink, and include solutions, suspensions, emulsions, powders, and solid moldings. It is not particularly limited as long as it can be taken orally such as food.
  • foods and drinks and food and drink compositions include milk products (including processed milk), yogurts, lactic acid bacteria beverages, fermented milk, ice creams, creams, cheeses, and other dairy products;
  • milk products including processed milk
  • yogurts lactic acid bacteria beverages
  • fermented milk ice creams, creams, cheeses, and other dairy products
  • fruit juice beverages vegetable beverages, soy milk beverages, coffee beverages, tea beverages, jelly beverages, cocoa, smoothie powdered beverages and sports powdered beverages
  • fortified powdered beverages cosmetic powdered foods, powdered soups, steamed bread
  • beverages such as concentrated beverages and alcoholic beverages
  • bread products such as bread, pasta, noodles, cake mixes, fried flour and bread crumbs
  • chocolate, gum, rice cake, cookies, gummi, snacks Japanese sweets, jelly, pudding, etc.
  • Confectionery such as dessert confectionery; curry, busta sauce, potov, stew, Japanese food retort food; processed oil, butter, margarine, spread Fats and oils such as mayonnaise; Instant foods such as freeze-dried foods; Agricultural processed products such as canned agricultural products, jams and marmalades, pickles, boiled beans, cereals, miscellaneous foods; processed fishery products; processed livestock products; Frozen foods such as side dishes and fries; liquid foods, animal feeds, tablets, and cosmetics used in the oral cavity.
  • the food and drink and the food and drink composition include functional food, health and nutrition food, health food, food for specified health, functional display food, functional nutrition food, food for the sick, Also included are classifications such as infant formula, maternal or lactating infant formula, or food and drink with a disease risk reduction label.
  • the indication of disease risk reduction is the indication of food or drink that may reduce the disease risk, and is based on the standard established by the FAO / WHO Joint Food Standards Committee (Codex Committee) or Refers to the standard and is a prescribed or recognized indication.
  • arbitrary components can be added to the food and drink and the food and drink composition as necessary.
  • optional ingredients are not particularly limited, but are usually sweeteners, acidulants, vegetables, fruits and seed juices and extracts, vitamins, minerals, amino acids, etc.
  • Nutrients lactic acid bacteria (excluding essential lactic acid bacteria according to embodiments of the present invention), useful microorganisms such as bifidobacteria and propionic acid bacteria, fermented products thereof, functional sugars such as oligosaccharides, royal jelly, glucosamine , Existing functional materials such as astaxanthin and polyphenol, fragrances, pH adjusters, excipients, acidulants, colorants, emulsifiers, preservatives and the like.
  • composition for preserving DNA is preferably taken orally for at least 7 days so that the intake of the polysaccharide is 200 ⁇ g or more / day.
  • the intake of the polysaccharide is not particularly limited as long as it does not harm the human body, but considering the cost effectiveness, it is preferably in the range of 200 ⁇ g / day to 60000 ⁇ g / day, preferably 300 ⁇ g / day to 45000 ⁇ g. Is more preferably in the range of 400 ⁇ g / day to 30000 ⁇ g / day, and particularly preferably in the range of 500 ⁇ g / day to 15000 ⁇ g / day. And “mass / day or more” is synonymous with “mass / day / day”, and “mass / day or less” is synonymous with “mass / day / day”.)
  • the required doses of the above-mentioned active ingredients are based on human administration experiments, but the necessary doses to the human body from the required administration doses in animal experiments (for example, mouse experiments) using the following formula based on Food Safety Commission materials. It can also be converted into a dose.
  • (Necessary administration dose to human body (converted value)) (Necessary administration dose to animal) ⁇ (Lower female body weight: 40 kg) ⁇ (Safety factor: 100)
  • the content of polysaccharides in yogurt was measured according to the phenol sulfate method (Hodge et al., “Methods in carbohydrate chemistry”, Vol. 1, page 338 (1962)). Specifically, it is as follows.
  • the yogurt was administered at 11.3 g / kg body weight / day, and the polysaccharide extract group was administered at 70 mg / kg body weight / day.
  • the dorsal skin of hairless mice in the pre-UV irradiation water group, the pre-UV irradiation yogurt group, and the pre-UV irradiation polysaccharide extract group was collected under isoflurane anesthesia. Further, after 7 days from the start of administration of each sample, the back of the remaining group of hairless mice was irradiated with ultraviolet light at an illuminance of 0.4 mW / cm 2 for 50 seconds (GL20SE manufactured by Sankyo Electric Co., Ltd.
  • a group of hairless mice administered with water (corresponding to a comparative example) is referred to as a “control group”, and a group of hairless mice orally administered with yogurt at 11.3 g / kg body weight / day (in the examples).
  • polysaccharide extract group a group of hairless mice (corresponding to Examples) in which the polysaccharide extract was orally administered at 70 mg / kg body weight / day is referred to as “polysaccharide extract group”.
  • DNA damage marker Verification of inhibitory effect of DNA damage by active oxygen DNA was collected from the back skin of each group of hairless mice collected as described above using qiagen QIAamp DNA Mini Kit (Qiagen) and purified. . Then, this DNA was degraded using DNase I, alkaline phosphatase, and phosphodiesterase (8-OHdG Assay Preparation Reagent Set, manufactured by Wako Pure Chemical Industries, Ltd.). Thereafter, the abundance of 8-oxo-2′-deoxyguanosine (8-OHdG), a DNA damage marker, was quantified.
  • a high performance liquid chromatography / tandem mass spectrometer (4500QTRAP, manufactured by Sciex) was used.
  • ACQUITY UPLC BEH C18 (1.7 ⁇ m, 2.1 mm ⁇ 100 mm, manufactured by Waters) was used.
  • 0.1% formic acid aqueous solution was used as mobile phase A
  • 0.1% formic acid methanol solution was used as mobile phase B.
  • the mobile phase composition at the start was set to mobile phase A: 100%, and after 5 minutes, the mobile phase composition was “mobile phase A: 50%, mobile phase B: 50%”.
  • the mobile phase composition was set to “mobile phase A: 1%, mobile phase B: 99%” and held for 1 minute, and the mobile phase composition was switched to mobile phase A: 100% and held for 2 minutes. did.
  • the measurement time of one sample in the high performance liquid chromatography / tandem mass spectrometer is set to 8 minutes, the flow rate of the mobile phase is set to 0.3 mL / min, the column temperature is set to 40 ° C., and electrospray ionization is used. 8-OHdG was detected in positive mode.
  • the analytical variables of the high performance liquid chromatography / tandem mass spectrometer were set as follows.
  • RNA was extracted from the back skin of each group of hairless mice collected as described above using RNeasy Mini Kit (Qiagen). Thereafter, cDNA was synthesized from the extracted total RNA using a RiverAid First Strand cDNA Synthesis Kit (manufactured by Thermo Fisher). Then, using ABI 7500 Fast Realtime PCR system (Applied Biosystems), the relative amount of the catalase mRNA was quantified from the cDNA and GAPDH as the endogenous gene.
  • XPA xeroda pigmentmentum complementation group A
  • the results of this experimental example are as shown in FIG. 4.
  • the relative amount of XPA mRNA which is a DNA repair protein after 24 hours and 72 hours after UV irradiation, is higher than that of the control group in the yogurt group and polysaccharide extract.
  • yogurt and polysaccharide extract were found to have an effect of promoting the repair of DNA damage caused by ultraviolet irradiation or active oxygen.
  • the symbol “*” indicates that there is a significant difference with respect to the control group at P ⁇ 0.05.

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Abstract

La présente invention vise à fournir une composition destinée à conserver de l'acide désoxyribonucléique qui garantit une quantité d'approvisionnement suffisante et contribue à une réduction du coût de production. Selon la présente invention, la composition destinée à la conservation d'acide désoxyribonucléique comprend, en tant que principe actif, un produit comportant un polysaccharide de bactéries lactiques. Le produit de bactéries lactiques est, de préférence, produit par l'utilisation combinée de la souche Lactobacillus delbrueckii subsp. bulgaricus OLL1247 avec la souche Streptococcus thermophilus OLS3078.
PCT/JP2017/025335 2016-07-19 2017-07-12 Composition destinée à la conservation d'acide désoxyribonucléique et procédé de production de ladite composition, procédé de conservation d'acide désoxyribonucléique, produit de bactéries lactiques et procédé d'utilisation dudit produit de bactéries lactiques WO2018016393A1 (fr)

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