WO2011025019A1 - Bactérie lactique présentant une activité induisant la thiorédoxine et aliments, boissons et médicaments pour prévenir et/ou améliorer un dommage biologique par l'intermédiaire de la thioredoxine - Google Patents

Bactérie lactique présentant une activité induisant la thiorédoxine et aliments, boissons et médicaments pour prévenir et/ou améliorer un dommage biologique par l'intermédiaire de la thioredoxine Download PDF

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Publication number
WO2011025019A1
WO2011025019A1 PCT/JP2010/064756 JP2010064756W WO2011025019A1 WO 2011025019 A1 WO2011025019 A1 WO 2011025019A1 JP 2010064756 W JP2010064756 W JP 2010064756W WO 2011025019 A1 WO2011025019 A1 WO 2011025019A1
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Prior art keywords
lactic acid
thioredoxin
acid bacterium
present
product
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PCT/JP2010/064756
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English (en)
Japanese (ja)
Inventor
俊彦 大澤
裕之 伊藤
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明治乳業株式会社
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Priority to JP2011528901A priority Critical patent/JP5851242B2/ja
Publication of WO2011025019A1 publication Critical patent/WO2011025019A1/fr

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    • 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
    • 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
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P39/00—General protective or antinoxious agents
    • A61P39/06—Free radical scavengers or antioxidants
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C12N1/205—Bacterial isolates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/225—Lactobacillus

Definitions

  • the present invention relates to a lactic acid bacterium having a thioredoxin-inducing activity and a method for using the lactic acid bacterium, and to a food and drink and a pharmaceutical product containing the lactic acid bacterium and used for the prevention and / or improvement of biological injury via thioredoxin.
  • thioredoxin is an in vivo substance that has been attracting attention in recent years.
  • Thioredoxin is known as a protein having two cystine residues in the active site and capable of regulating its activity through redox of an enzyme disulfide bond.
  • various oxidative stresses can be removed by the oxidation-reduction reaction, thereby preventing biological damage (for example, Non-Patent Documents 1, 2, and 3).
  • thioredoxin can be expressed and used effectively in vivo, it can prevent and prevent oxidative stress and thus prevent various injuries.
  • thioredoxin inducer containing an aldehyde compound having an ⁇ , ⁇ -unsaturated bond (Patent Document 1), a sulforaphane derived from a cruciferous plant (Patent Document 2), mugwort, mugwort extract, blue Examples include diso and blue diso extracts (Patent Document 3), isoprenoid compounds, geranylgeranylacetone (GGA) (Patent Document 4), and the like.
  • An object of the present invention is to provide a microorganism having thioredoxin-inducing activity.
  • the present inventors have conducted extensive research and found a lactic acid bacterium having a thioredoxin-inducing activity to complete the present invention.
  • the present invention relates to a lactic acid bacterium suitable for foods and beverages and pharmaceuticals, capable of inducing thioredoxin and preventing and / or improving various biological injuries caused by oxidative stress. Furthermore, this invention relates to the food-drinks and pharmaceutical which prevent and / or improve the biological injury using the said lactic acid bacteria.
  • one embodiment of the present invention is a lactic acid bacterium belonging to the genus Lactobacillus having thioredoxin-inducing activity.
  • the lactic acid bacterium is preferably a bacterium belonging to the genus Lactobacillus bulgaricus, more preferably Lactobacillus delbruecki subspecies bulgaricus (Lactobacillus delbrueckii subsp.
  • another aspect of the present invention is a food or drink for preventing and / or improving biological injury through thioredoxin, comprising the lactic acid bacterium of the present invention, or a lactic acid bacterium-containing product and / or a lactic acid bacterium-treated product.
  • Still another embodiment of the present invention is a food or beverage composition for preventing and / or improving biological injury through thioredoxin, comprising the lactic acid bacterium of the present invention, or a lactic acid bacterium-containing product and / or a processed product of lactic acid bacterium. .
  • Still another embodiment of the present invention is a pharmaceutical product for preventing and / or improving thioredoxin-mediated biological injury comprising the lactic acid bacterium of the present invention, or a lactic acid bacterium-containing product and / or a lactic acid bacterium-treated product.
  • lactic acid bacteria having thioredoxin-inducing activity can induce thioredoxin in vivo.
  • the lactic acid bacteria according to the present invention can be provided in various forms such as bacteria-containing substances and bacteria-treated substances. It can be provided as a food / beverage product, a food / beverage product composition, and a pharmaceutical that prevent and / or improve various biological injuries.
  • lactic acid bacteria having thioredoxin-inducing activity of the present invention include fungi belonging to the genus Lactobacillus, such as Lactobacillus gasseri strain, Lactobacillus bulgaricus strain, and the like. Can do.
  • the present invention is not limited to these species, and these strains can be used alone or in combination of two or more.
  • the Lactobacillus bulgaricus strain is particularly preferable. More preferred is Lactobacillus delbrueckii subsp. Bulgaricus OLL1224 strain.
  • Lactobacillus delbrueckii subsp. Bulgaricus OLL1224 strain used in the present invention has been deposited under the following conditions and deposited under the Budapest Treaty.
  • Bulgaricus OLL1224 strain is a plant-derived Gram-positive obligate anaerobic gonococcus.
  • Lactobacilli MRS ro Broth (BD) is inoculated with this bacterium and cultured at 37 ° C for 18 hours under anaerobic conditions using AnaeroPack Kenki (Mitsubishi Gas Chemical Co., Ltd.). Type), flat (partially conical) colonies are observed.
  • the lactic acid bacterium having thioredoxin-inducing activity of the present invention may be prepared as a substance containing the lactic acid bacterium or a processed product thereof.
  • the content of the present invention or the processed product thereof is not limited to the following, and specifically, a culture, concentrate, pasted product, spray-dried product, freeze-dried product, vacuum-dried product of lactic acid bacteria having thioredoxin-inducing activity.
  • the lactic acid bacterium having thioredoxin-inducing activity of the present invention can induce or increase the expression of thioredoxin-related genes including thioredoxin genes.
  • This lactic acid bacterium of the present invention can prevent or reduce biological damage, preferably various biological damages caused by oxidative stress, by inducing or increasing the expression of thioredoxin.
  • the present invention comprises administering a lactic acid bacterium having thioredoxin-inducing activity of the present invention, a content or processed product of the lactic acid bacterium, or a pharmaceutical composition (medicine) containing them to an animal (which may or may not include humans).
  • a lactic acid bacterium having thioredoxin-inducing activity of the present invention a content or processed product of the lactic acid bacterium, or a pharmaceutical composition (medicine) containing them to an animal (which may or may not include humans).
  • a pharmaceutical composition containing them to an animal (which may or may not include humans).
  • the present invention also relates to a lactic acid bacterium having thioredoxin-inducing activity of the present invention, a content or treated product of the lactic acid bacterium, or a pharmaceutical composition (pharmaceutical product) containing them for use in the prevention and / or improvement of biological injury.
  • a lactic acid bacterium having thioredoxin-inducing activity of the present invention a content or treated product of the lactic acid bacterium, or a pharmaceutical composition (pharmaceutical product) containing them for use in the prevention and / or improvement of biological injury.
  • the intake or dose of lactic acid bacteria having thioredoxin-inducing activity of the present invention can be appropriately set in consideration of various factors such as the route of administration, the age, weight, and symptoms of animals to be administered including humans.
  • the dose of the lactic acid bacteria is preferably 1 ⁇ 10 5 to 1 ⁇ 10 10 cfu / g, more preferably 1 ⁇ 10 6 to 1 ⁇ 10 9 cfu / g per adult day. Ingest or administer.
  • the amount may be smaller than the above range, and since this active ingredient has no safety problem, Can be used in large quantities.
  • the lactic acid bacteria having thioredoxin-inducing activity of the present invention are preferably administered orally.
  • the lactic acid bacterium having thioredoxin-inducing activity of the present invention can be administered (preferably orally administered) in the form of a lactic acid bacterium-containing product or processed product, or a pharmaceutical composition (pharmaceutical product) containing them.
  • the lactic acid bacterium having thioredoxin-inducing activity of the present invention, the lactic acid bacterium-containing product and the processed product thereof can be used as a pharmaceutical or a pharmaceutical composition for preventing and / or improving various biological injuries caused by oxidative stress, or for its production. it can.
  • the administration form of pharmaceuticals include oral administration by tablets, coated tablets, capsules, granules, powders, solutions, syrups, emulsions and the like.
  • lactic acid bacteria having thioredoxin-inducing activity are prepared by adding lactic acid bacteria having thioredoxin-inducing activity according to the present invention, the lactic acid bacteria-containing product and / or processed products thereof to excipients, binders, disintegrating agents, lubricants, coloring agents, flavoring agents, and the like. It can be formulated using known adjuvants that can be generally used in the pharmaceutical formulation technical field, such as agents, solubilizers, suspensions, and coating agents.
  • the lactic acid bacterium having thioredoxin-inducing activity of the present invention, the lactic acid bacterium-containing product, and the treated product thereof can be used for foods and beverages that prevent and / or improve various biological injuries caused by oxidative stress, or the production thereof.
  • the food and drink of the present invention are not limited in category and type, and may be functional foods, foods for specified health use, health foods, foods for nursing care, dairy products such as confectionery, lactic acid bacteria beverages, cheese and yogurt, and seasonings Etc.
  • dairy products such as confectionery, lactic acid bacteria beverages, cheese and yogurt, and seasonings Etc.
  • the above food and drink can be produced by ordinary methods of those skilled in the art.
  • lactic acid bacteria having thioredoxin-inducing activity of the present invention can be processed into general foods and drinks including dairy products and fermented milk as described above, as well as for producing dairy products and fermented milk such as yogurt and cheese. It can also be used as a starter. In the case of a starter, other microorganisms may be mixed as long as there is no hindrance to the inhabiting and growth of Lactobacillus lactic acid bacteria of the present invention, and there is no hindrance to dairy production.
  • the starter may be mixed with Streptococcus thermophilus strain, Lactobacillus acidophilus strain, which is the main bacterial species as lactic acid bacteria for yogurt, and other bacterial species generally used for yogurt and cheese Can be used as a starter.
  • Streptococcus thermophilus strain Lactobacillus acidophilus strain, which is the main bacterial species as lactic acid bacteria for yogurt, and other bacterial species generally used for yogurt and cheese Can be used as a starter.
  • Production of dairy products and fermented milk using the starter can be performed according to a conventional method.
  • plain yogurt can be produced by mixing the above starter with milk or a dairy product cooled after heating, mixing, homogenization, and sterilization, and then fermenting and cooling.
  • the food and drink of the present invention specifically includes various foods and drinks, food and drink compositions (for example, milk, soft drinks, fermented milk, yogurt, cheese, bread, biscuits, crackers, pizza crusts, prepared milk powder, and liquid foods.
  • the state of the food / beverage products used normally, for example, any of solid (powder, granule, etc.), paste, gel, liquid or suspension may be sufficient.
  • lactic acid bacterium-containing product and / or the lactic acid bacterium-treated product in a food or drink or a food or drink composition water, protein Sugars, lipids, vitamins, minerals, organic acids, organic bases, fruit juices, flavors and the like can be used as main components.
  • the protein examples include whole milk powder, skim milk powder, partially skim milk powder, casein, whey powder, whey protein, whey protein concentrate, whey protein isolate, ⁇ -casein, ⁇ -casein, ⁇ -casein, ⁇ -lactoglobulin , ⁇ -lactalbumin, lactoferrin, soy protein, chicken egg protein, meat protein and other animal and vegetable proteins, their hydrolysates, butter, whey minerals, cream, whey, non-protein nitrogen, sialic acid, phospholipids, lactose, etc. And various milk-derived components.
  • saccharide examples include general saccharides, modified starch (in addition to dextrin, soluble starch, British starch, oxidized starch, starch ester, starch ether, etc.), dietary fiber, and the like.
  • lipid examples include animal oils such as lard, fish oil, etc., fractionated oils, hydrogenated oil, transesterified oil, etc .; palm oil, safflower oil, corn oil, rapeseed oil, coconut oil, fractionated oils thereof, Examples include vegetable oils such as hydrogenated oils and transesterified oils.
  • vitamins include vitamin A, carotene, vitamin B group, vitamin C, vitamin D group, vitamin E, vitamin K group, vitamin P, vitamin Q, niacin, nicotinic acid, pantothenic acid, biotin, inositol, choline.
  • minerals include, for example, calcium, potassium, magnesium, sodium, copper, iron, manganese, zinc, selenium, and whey minerals.
  • organic acid include malic acid, citric acid, lactic acid, and tartaric acid. These components can be used alone or in combination of two or more, and a synthetic product and / or a food containing a lot of these may be used.
  • the content of the lactic acid bacterium having thioredoxin-inducing activity, the lactic acid bacterium-containing product and / or the treated product of the present invention can be arbitrarily determined according to the purpose and application. Although the present invention is not limited to this, the content is preferably 1 ⁇ 10 5 to 1 ⁇ 10 10 cfu / g, more preferably 1 ⁇ 10 6 to 1 ⁇ 10 9 cfu / g.
  • Example 1 ⁇ Method for preparing methanol extract of lactic acid bacteria culture solution>
  • the lactic acid bacteria (Lactobacillus delbrueckii subsp. Bulgaricus) OLL1224 strain of the present invention was inoculated into 5 ml of TPYG medium shown in Table 1 at 37 ° C. for 18 hours in aneropack (Mitsubishi Gas Chemical).
  • the absorbance of the culture solution at 660 nm at the end of the culture was 4.530 (0.044 for the TPYG medium alone) After the culture, the culture solution was transferred to a stainless steel vat and freeze-dried.
  • the weight of the dried product was 0.61 g, and an equal amount of methanol to the culture solution before lyophilization was added to the lyophilized product, stirred well, and then extracted by allowing to stand at 5 ° C. for 40 hours. Then, stir well and collect the supernatant by centrifugation (12000G, 10 minutes) , which was used as a sample. Also, the medium without inoculation with lactic acid bacteria as a control (Control).
  • Example 2 ⁇ Array sample> Mouse fibroblast 3T3-L1 cells were seeded in 9 cm dishes. After confirming sufficient growth, the medium was replaced with a medium containing dexamethasone (1 ⁇ M), insulin (2 ⁇ M), and 3-isobutyl-1-methylxanthene (0.25 mM) to induce differentiation. Further, after 3, 5, and 7 days, the medium was replaced with a medium containing insulin (2 ⁇ M) to promote differentiation.
  • lactic acid bacterium OLL1224 strain of the present invention The cells were cultured in a medium (500 ⁇ L) supplemented with a sample containing% for 3 hours (experimental group).
  • the mouse fibroblast 3T3-L1 cells alone were used as the non-added group, and the group containing only the culture medium shown in Table 1 without the lactic acid bacteria culture of the present invention was used as the Control group.
  • the medium was discarded and washed twice with sterilized PBS, and then cultured for 20 hours in a medium containing TNF- ⁇ (10 ng / mL).
  • the supernatant was collected and appropriately diluted with PBS, and the amount of IL-6 was measured by ELISA.
  • the cells were washed twice with PBS and then dissolved in Trizol, and RNA for microarray samples was collected by a conventional method.
  • an antibody chip (Genopal, manufactured by Mitsubishi Rayon Co., Ltd.) carrying 221 types of genes involved in oxidative stress, inflammatory reaction, antioxidant enzyme, detoxification enzyme, or obesity was prepared and used to test the RNA recovered above. These gene expression data were obtained as samples.
  • the gene expression in the additive-free group was taken as 1, and the ratio of gene expression in the Control group and the experimental group was calculated.
  • Table 2 shows an example of an array sample.
  • Txndc2, Txnll1, and Txndc17 which are thioredoxin-related genes
  • the expression in the group to which the lactic acid bacteria culture of the present invention was added was higher than that in the Control group. Therefore, it was revealed that the lactic acid bacterium of the present invention has an activity of inducing thioredoxin.
  • Example 3 From the lactic acid bacteria confirmed to have thioredoxin-inducing activity in Example 2 above, Lactobacillus delbruecki subsp. Bulgaricus OLLL1224 strain, yogurt was produced as a lactic acid bacteria food and drink by a conventional method. Thus, it was possible to obtain a yogurt for preventing and / or ameliorating a biological injury via a desired thioredoxin containing the lactic acid bacterium of the present invention.
  • Example 4 From the lactic acid bacteria confirmed to have thioredoxin-inducing activity in Example 2 above, Lactobacillus delbrueckii subsp. Bulgaricus OLLL1224 strain, a lactic acid bacteria concentrate was prepared by a conventional method. To obtain a capsule for preventing and / or ameliorating a desired thioredoxin-containing biological injury containing the lactic acid bacterium of the present invention by producing a capsule using an appropriate pharmaceutical additive in combination with a conventional method. I was able to.
  • the lactic acid bacteria having thioredoxin-inducing activity By ingesting the lactic acid bacteria having thioredoxin-inducing activity according to the present invention, various biological injuries caused by oxidative stress can be prevented and / or improved. Moreover, since the lactic acid bacteria according to the present invention can be provided in various forms, they can be provided as foods and drinks or pharmaceuticals that can easily prevent and / or improve various biological injuries caused by oxidative stress. Since the technique using lactic acid bacteria is simple and there is no problem with respect to safety to the human body, the excellent effect can be easily and effectively utilized, and its utility value is high.

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Abstract

L'invention porte sur une bactérie lactique qui est appropriée pour des aliments, des boissons et des médicaments, qui induit de la thioredoxine et qui est capable de prévenir et/ou d'améliorer une diversité de dommages biologiques résultant d'un stress oxydant. L'invention porte également sur des aliments, des boissons et des médicaments utilisant ladite bactérie lactique qui empêche et/ou améliore un dommage biologique. La bactérie lactique présentant une activité induisant la thioredoxine appartient au genre Lactobacillus, en particulier à la souche Lactobacillus delbrueckii sous-espèce bulgaricus OLL1224.
PCT/JP2010/064756 2009-08-31 2010-08-31 Bactérie lactique présentant une activité induisant la thiorédoxine et aliments, boissons et médicaments pour prévenir et/ou améliorer un dommage biologique par l'intermédiaire de la thioredoxine WO2011025019A1 (fr)

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Application Number Priority Date Filing Date Title
JP2011528901A JP5851242B2 (ja) 2009-08-31 2010-08-31 チオレドキシン誘導活性を有する乳酸菌ならびにチオレドキシンを介する生体傷害の予防および／または改善用の飲食品および医薬品

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JP2009199157 2009-08-31
JP2009-199157 2009-08-31

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WO2011025019A1 true WO2011025019A1 (fr) 2011-03-03

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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH07228536A (ja) * 1994-02-16 1995-08-29 Meiji Milk Prod Co Ltd 乳酸菌を用いた免疫賦活剤
JPH107577A (ja) * 1996-06-17 1998-01-13 Yakult Honsha Co Ltd 血糖降下剤
WO2007114230A1 (fr) * 2006-03-30 2007-10-11 Kyoto University Agent favorisant la production de thioredoxine
JP2007269737A (ja) * 2006-03-31 2007-10-18 Morinaga Milk Ind Co Ltd インターロイキン産生調節剤、該インターロイキン産生調節剤を含む医薬組成物及び飲食品、並びにその製造方法
WO2007138961A1 (fr) * 2006-05-29 2007-12-06 Redox Bioscience Inc. Agent prophylactique ou thérapeutique convenant pour des troubles provoquéspar un facteur d'inhibition de migration de macrophages

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH07228536A (ja) * 1994-02-16 1995-08-29 Meiji Milk Prod Co Ltd 乳酸菌を用いた免疫賦活剤
JPH107577A (ja) * 1996-06-17 1998-01-13 Yakult Honsha Co Ltd 血糖降下剤
WO2007114230A1 (fr) * 2006-03-30 2007-10-11 Kyoto University Agent favorisant la production de thioredoxine
JP2007269737A (ja) * 2006-03-31 2007-10-18 Morinaga Milk Ind Co Ltd インターロイキン産生調節剤、該インターロイキン産生調節剤を含む医薬組成物及び飲食品、並びにその製造方法
WO2007138961A1 (fr) * 2006-05-29 2007-12-06 Redox Bioscience Inc. Agent prophylactique ou thérapeutique convenant pour des troubles provoquéspar un facteur d'inhibition de migration de macrophages

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
SHOGO TAMASHI ET AL: "Redox Seigyo Kinosei Shokuhin", SHUKAN IGAKU NO AYUMI, July 2005 (2005-07-01), pages 216 - 220 *
TAKATSUME Y. ET AL: "Enrichment of yeast thioredoxin by green tea extract through activation of Yapl transcription factor in Saccharomyces cerevisiae", J AGRIC FOOD CHEM., vol. 53, no. 2, 26 January 2005 (2005-01-26), pages 332 - 337 *
YOSHIFUMI TAKATSUME ET AL: "Thioredoxin Gan'yu Kinosei Shokuhin no Sosei o Mezashita Ryokucha Chushutsubutsu ni yoru Kobo Thioredoxin no Yudo Seisan, 30E0928", NIPPON NOGEI KAGAKUKAI TAIKAI KOEN YOSHISHU, vol. 2005, 2005, pages 273 *

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JP5851242B2 (ja) 2016-02-03

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