WO2011118807A1 - 酵母の培養方法 - Google Patents
酵母の培養方法 Download PDFInfo
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- WO2011118807A1 WO2011118807A1 PCT/JP2011/057446 JP2011057446W WO2011118807A1 WO 2011118807 A1 WO2011118807 A1 WO 2011118807A1 JP 2011057446 W JP2011057446 W JP 2011057446W WO 2011118807 A1 WO2011118807 A1 WO 2011118807A1
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- Prior art keywords
- yeast
- culture
- citric acid
- liquid medium
- culturing
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- 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
- C12P21/00—Preparation of peptides or proteins
- C12P21/02—Preparation of peptides or proteins having a known sequence of two or more amino acids, e.g. glutathione
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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
- A23L27/00—Spices; Flavouring agents or condiments; Artificial sweetening agents; Table salts; Dietetic salt substitutes; Preparation or treatment thereof
- A23L27/20—Synthetic spices, flavouring agents or condiments
- A23L27/24—Synthetic spices, flavouring agents or condiments prepared by fermentation
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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
- A23L33/14—Yeasts or derivatives thereof
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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
- A23L33/14—Yeasts or derivatives thereof
- A23L33/145—Extracts
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- 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
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Definitions
- the present invention relates to a yeast culture method capable of increasing glutathione content in bacterial cells, and a method for producing a yeast extract from yeast cultured by the culture method.
- Yeasts belonging to the genus Saccharomyces such as brewer's yeast and baker's yeast, contain natural vitamin B groups, amino acids, minerals, etc. in a well-balanced manner. It is effectively used.
- dry yeast has been used in Japan for many years as a pharmaceutical, a raw material for food, and a seasoning, and is recognized as a highly nutritious and safe material. In recent years, it has also been widely used as a raw material yeast for yeast extract.
- Yeast extract is prepared from a yeast culture and contains abundant amino acids and has been used as a food additive such as a seasoning for imparting umami and richness.
- a seasoning for imparting umami and richness.
- the demand for yeast extract as a seasoning is increasing.
- Yeast extract prepared from yeast containing abundant taste components can be expected to be used as a better seasoning, and therefore, development of yeasts containing more taste components has been actively conducted.
- Examples of typical sulfur-containing compounds in yeast cells include glutathione and S-adenosylmethionine.
- Glutathione is a very useful substance having liver function recovery, antioxidant activity, and the like.
- glutathione is expected to be used in a wide range of applications such as seasonings, health foods and other food additives, and cosmetic base materials.
- S-adenosylmethionine is known to act as a methyl group donor in various biological reactions.
- effects such as antidepressant action, arthropathy alleviation, and liver function recovery have been reported, and it is known that these sulfur-containing compounds play an important role for the living body.
- the sulfur-containing compound is usually synthesized by transcription and translation products of many genes including a MET gene (methionine synthesis gene) group using sulfur-containing amino acids such as methionine and cysteine. Therefore, in order to obtain a yeast that produces a higher amount of sulfur-containing compounds, a mutation is caused in the genes related to the synthesis of these sulfur-containing compounds that the yeast has, and a yeast mutant containing a high sulfur-containing compound content is produced.
- a method for producing a yeast having a high glutathione content (1) aerobically cultivating a mutant strain of Candida yeast that has become capable of growing in a medium containing ethionine and sulfite by mutation treatment.
- a method for increasing the glutathione content in the bacterial cells has been disclosed (see, for example, Patent Document 1).
- a yeast having a high glutathione content can also be obtained by a method of screening a yeast having a higher glutathione content from yeasts that have been genetically modified by mutation treatment or the like.
- the mutation treatment and screening require labor and labor, and it is often not always possible to obtain a glutathione-rich yeast.
- natural yeast wild strain
- development of a method for increasing glutathione content in yeast without mutation treatment is desired. For example, when yeast or yeast extract is used for food, a wild strain may be preferred over a recombinant.
- An object of the present invention is to provide a method for culturing yeast that can increase the glutathione content in cells without genetic modification.
- the present inventors have added a predetermined amount or more of citric acid to a liquid medium when cultivating yeast such as Saccharomyces sp.
- the present invention was completed.
- the present invention (1) A method for culturing yeast, comprising culturing yeast in a liquid medium having a citric acid concentration of 20 mM or more, (2) The yeast culture method according to (1), wherein the liquid medium has a citric acid concentration of 200 mM or less, (3) The method for culturing yeast according to (1) or (2) above, wherein the citrate concentration of the liquid medium at the start of the culture is 20 mM or more, (4) including adjusting the citrate concentration of the liquid medium to 20 to 200 mM when the citrate concentration of the liquid medium at the start of the culture is less than 20 mM and the growth state of the yeast is in the induction phase or the logarithmic growth phase.
- the yeast culture method according to (1), (5) The citrate concentration of the liquid medium at the start of the culture is less than 20 mM, and the citrate concentration of the liquid medium is adjusted to 20 to 200 mM within 9 hours after the start of the culture (1) The yeast culture method described. (6) The yeast culture method according to any one of (1) to (5), wherein the yeast is a genus Saccharomyces or a genus Candida, (7) The yeast culture method according to any one of (1) to (5), wherein the yeast is Saccharomyces cerevisiae or Candida utilis.
- a method for increasing the glutathione content of yeast comprising culturing yeast in a liquid medium having a citric acid concentration of 20 mM or more
- a method for producing yeast comprising recovering yeast cultured by the yeast cultivation method according to any one of (1) to (7) above
- a method for producing a yeast extract comprising extracting a yeast extract from a yeast cultured by the yeast culturing method according to any one of (1) to (7), (11)
- a method for producing a food or drink including use as a raw material, (12) A yeast extract prepared from a yeast cultured by the yeast culture method according to any one of (1) to (7) above, (13) A seasoning composition containing the yeast cultured by the yeast culturing method according to any one of (1) to (7) or the yeast extract according to (12), (14) Yeast cultivated by the yeast culturing method according to any one of (1) to (7), or a food or drink containing the yeast extract according to (12), Is to provide.
- the glutathione content of yeast such as Saccharomyces can be increased by a simple process of culturing in a liquid medium containing a sufficient amount of citric acid. Moreover, since the yeast cultivated by the culture method has a sufficiently high glutathione content, by using the yeast, yeast extracts and foods and drinks having a high glutathione content can be easily obtained.
- Example 1 it is the graph which showed GSH content rate (%) for every citric-acid density
- Example 2 it is the graph which showed the GSH content rate (%) of each culture for every citric acid concentration added to molasses culture medium (1).
- Example 2 it is the graph which showed pH (final pH) of the liquid culture medium at the time of completion
- the Saccharomyces cerevisiae KK122 strain is a view showing a measurement result of the GSH content and OD 600.
- Example 5 the Saccharomyces cerevisiae KK124 strain is a view showing a measurement result of the GSH content and OD 600.
- Example 11 it is the figure which showed the result of having measured the GSH content rate (%) of the yeast in each sample.
- the reference example 1 it is the figure which showed the measurement result of the citric acid content in each supernatant before and behind culture
- glutathione means both oxidized glutathione and reduced glutathione
- the total glutathione content means the total content of oxidized glutathione and reduced glutathione.
- the glutathione content per dry cell weight of yeast can be determined by a method usually performed when the glutathione content in a microorganism is quantified.
- the total glutathione content per dry cell weight of yeast can be measured according to the method of Titze et al. (Analytical Biochemistry, Vol. 27, p502, 1969).
- DTNB 5,5′-dithiobis (2nitrobenzoic acid)
- NADPH nicotinamide adenine dinucleotide phosphate reduced form
- the reaction rate of the reaction is proportional to the amount of glutathione present. This is a method for measuring glutathione content.
- the yeast culture method of the present invention is characterized in that yeast is cultured in a liquid medium having a citric acid concentration of 20 mM or more.
- yeast is cultured in a liquid medium having a citric acid concentration of 20 mM or more.
- the glutathione content of the yeast can be increased, and yeast with a high glutathione content (yeast with a high glutathione content) can be obtained.
- the reason why such an effect of high glutathione content (effect of increasing the glutathione content of yeast) is not clear.
- Example 3 described later when other acids are added to the liquid medium, the effect is not observed, and the effect is obtained by adding citric acid even in a pH controlled environment. Therefore, it is not just the pH adjustment effect of the liquid medium, but the production of glutathione is promoted by some action specific to citric acid, or the discharge to the outside of the cell is suppressed, so that It is presumed that glutathione is accumulated.
- One aspect of the yeast culturing method of the present invention is a method for producing a glutathione-rich yeast, wherein the yeast is cultured in a liquid medium having a citric acid concentration of 20 mM or more. And a method for producing a yeast having a high glutathione content, which comprises recovering the yeast from the culture.
- the glutathione-rich yeast of the present invention refers to a yeast in which the glutathione content in the microbial cells is significantly increased compared to the parent strain.
- the culture means a culture containing yeast cells and a medium used for culturing yeast.
- the yeast used in the yeast culture method of the present invention is not particularly limited, but is preferably Saccharomyces sp. Or Candida sp.
- Saccharomyces cerevisiae Saccharomyces cerevisiae
- Saccharomyces paradoxus Saccharomyces paradoxus
- Saccharomyces Mikatae Saccharomyces mikatae
- Saccharomyces bayanus Saccharomyces bayanus
- Saccharomyces click Doria Vu zero Vii Sacharomyces kudriavzevii
- Candida utilis Candida utilis
- Candida tropicalis Candida lipolytica
- Candida salmon etc.
- the yeast culturing method of the present invention can provide a high glutathione-containing effect not only when cultivating wild strains (natural yeast) but also when cultivating mutant strains obtained by mutation treatment.
- wild strain means a yeast that originally existed in nature, that is, a yeast that has not been subjected to artificial mutation treatment.
- mutant strain means a yeast obtained by subjecting a gene to artificial mutation treatment.
- the mutation treatment is not particularly limited as long as it is a treatment capable of mutating a part of a gene possessed by an organism such as yeast, and when producing a mutant strain of a microorganism such as yeast. Any commonly used technique may be used.
- the yeast can be mutated by treating the yeast with ultraviolet rays, ionizing radiation, nitrous acid, nitrosoguanidine, ethyl methanesulfonate (hereinafter abbreviated as EMS) or the like as a mutagen. .
- the citric acid concentration of the liquid medium used in the yeast culture method of the present invention may be 20 mM or more.
- the citric acid concentration is 20 mM or more, a sufficient amount of citric acid can be allowed to act on the yeast in order to achieve the effect of high glutathione content.
- the citrate concentration of the liquid medium is preferably 20 to 200 mM, more preferably 20 to 120 mM, further preferably 20 to 100 mM, and particularly preferably 50 to 100 mM. preferable.
- an excessive amount of citric acid is added to the liquid medium, the effect of high glutathione content cannot be obtained as expected, and there is a risk of inhibiting the growth of the yeast.
- By setting the amount to 200 mM or less, a sufficient glutathione high content effect can be obtained while suppressing the influence on the growth of yeast.
- the glutathione content is slightly higher compared to the liquid medium not added even when an acid other than citric acid is added. Become. This is presumably because the pH of the liquid medium was adjusted by the buffering action due to acid addition. That is, when the pH is not controlled, a part of the citric acid added to the liquid medium is used for pH control of the liquid medium. For this reason, the citric acid concentration of the liquid medium necessary for obtaining the same high glutathione content effect tends to be higher in the culture conditions where the pH is not controlled than in the culture conditions where the pH is controlled.
- the yeast culturing method of the present invention it is preferable to culture the yeast at a citric acid concentration of 60 to 110 mM, and culture the yeast at 75 to 90 mM. More preferably.
- the yeast culture method of the present invention the yeast is cultured at a citrate concentration of 20 to 100 mM in the liquid medium. It is preferable to culture yeast at 20 to 75 mM.
- the yeast may be cultured using a liquid medium adjusted in advance so that the citric acid concentration is 20 mM or more, and citric acid is added to the liquid medium after the start of the culture. May be. That is, the yeast may be cultured with a citric acid concentration of 20 mM or more in the liquid medium at the start of the culture, and the citric acid concentration of the liquid medium is less than 20 mM (including a liquid medium containing no citric acid at the start). After culturing the yeast, the citric acid concentration of the liquid medium may be adjusted to 20 to 200 mM after the start of the culture. In the method of adjusting the citric acid concentration of the liquid medium, solid citric acid may be added to the liquid medium for adjustment, or an aqueous citric acid solution may be added for adjustment.
- the earlier the citric acid is added to the liquid medium the higher the glutathione content tends to be obtained.
- the effect of high glutathione content is sufficiently exerted by allowing a sufficient concentration of citric acid to act on yeast in a state of citizenship. Therefore, when adjusting the citrate concentration of the liquid medium after the start of culture, before entering the stationary phase, i.e., the yeast growth state is in the induction phase or logarithmic growth phase, preferably in the logarithmic growth phase after the start of the culture.
- the logarithmic growth phase is a period in which, in batch and fed-batch culture, when the amount of yeast in the culture vessel is measured over time using absorbance or the like as an index, it is observed that the logarithmic growth period increases logarithmically.
- continuous culture it is a time when the amount of yeast in the culture vessel is observed to be almost constant.
- the induction period is the period from the start of culture to the logarithmic growth phase in batch and fed-batch culture.
- continuous culture it is the time until the parameter that becomes Merckmar is controlled to a constant value. For example, by adjusting the citric acid concentration of the liquid medium within 9 hours, preferably within 3 hours from the start of culture, a sufficient concentration of citric acid can be allowed to act on yeast in a state where budding is thriving.
- liquid medium used in the yeast culturing method of the present invention a liquid medium in which yeast can grow is appropriately added with citric acid so that the citric acid concentration becomes 20 mM or more.
- the liquid medium to which citric acid is added contains a carbon source, a nitrogen source, an inorganic salt, and the like, and any medium that is usually used for culturing yeasts such as Saccharomyces cerevisiae can be used. .
- the carbon source contained in the liquid medium is, for example, one or more selected from the group consisting of glucose, sucrose, acetic acid, ethanol, molasses, sulfite pulp waste liquid, and the like that are used for normal microorganism culture Substances can be used.
- the nitrogen source may be a nitrogen-containing inorganic salt or a nitrogen-containing organic substance.
- one or more substances selected from the group consisting of urea, ammonia, ammonium sulfate, ammonium chloride, ammonium phosphate, corn steep liquor (CSL), casein, yeast extract, and peptone can be used. .
- inorganic salts phosphoric acid components such as lime superphosphate and phosphorous acid, potassium components such as potassium chloride and potassium hydroxide, and magnesium components such as magnesium sulfate and magnesium hydrochloride can be used.
- inorganic salts such as zinc, copper, manganese, and iron ions may be used.
- vitamins, nucleic acid-related substances and the like may be appropriately added to the liquid medium used in the yeast culture method of the present invention.
- the liquid medium used in the yeast culture method of the present invention may be a synthetic medium such as an SD medium, or a semi-synthetic medium such as a YPD medium or a molasses medium.
- a medium originally containing a small amount of citric acid such as a molasses medium, may be used.
- the culture medium which modified these may be sufficient.
- molasses medium sucrose concentration 3%) varies in content depending on the lot, etc.
- the total organic acid concentration is generally about 1 to 3.5 g / L
- the citric acid concentration is 50 to 50%. 400 mg / L. That is, it is calculated from the molecular weight 192 of citric acid that the molasses medium used for yeast culture usually contains 0.2 to 2 mM citric acid.
- citric acid may be used as a carbon source when culturing yeast.
- citric acid affects the pH of the medium
- citric acid is actively added to the medium (for example, citric acid is added so as to be 10 mM or more).
- citric acid added to the liquid medium is used as a carbon source. It is clear that it is not used.
- the culture format is not particularly limited as long as it uses a liquid medium, and can be appropriately determined in consideration of the culture scale and the intended use of the obtained culture.
- Examples of the culture format in the liquid medium include batch culture, fed-batch culture, and continuous culture.
- the culture conditions of the yeast culture method of the present invention are not particularly limited, except that a liquid medium containing a predetermined concentration of citric acid is used, and culture is performed under conditions generally used for culturing yeast. Can do.
- the culture temperature is preferably 20 to 40 ° C, more preferably 25 to 35 ° C.
- the pH of the medium is preferably 3.5 to 8.0, and more preferably 4.0 to 6.0. In particular, when industrially mass-producing a culture, it is preferable to periodically measure the pH in the medium and adjust the pH to 4.0 to 6.0.
- the amount of aeration and the conditions for stirring can be appropriately determined in consideration of the culture volume and time, and the initial concentration of bacteria.
- the ventilation is 0.2-2V.
- V. M.M. Stirring can be performed at about (Volume per volume per minuts) and about 50 to 800 rpm.
- the yeast having a high glutathione content can be cultured by the yeast culturing method of the present invention.
- the dried yeast cells are more cultivated than in the case of culturing in a liquid medium having a citric acid concentration of less than 20 mM (including a liquid medium containing no citric acid).
- the glutathione content contained therein can be increased by 10% or more.
- dry cell weight means the weight after drying a cell.
- the yeast culture is centrifuged to collect the microbial cells as a precipitate.
- the weight of the dried cells can be determined by washing the collected cells twice with a centrifugal separation and then measuring the weight after drying at 105 ° C. for 5 hours.
- the yeast culture method of the present invention including culturing yeast in a liquid medium having a citric acid concentration of 20 mM or more, a culture containing yeast having a high glutathione content can be obtained.
- the yeast can be recovered from the product.
- the method for recovering the yeast from the culture is not particularly limited, and any of the recovery methods that are usually performed when recovering the yeast from the yeast culture may be used. Examples of a method for recovering yeast from the culture include a method of centrifuging a yeast culture.
- the yeast extract of this invention can be prepared by extracting a yeast extract from the yeast cultured with the yeast culture
- glutathione is a substance having various physiological activities. That is, it is possible to produce a yeast or yeast extract with higher added value than conventional by simply applying the yeast culture method of the present invention instead of the conventional culture method.
- Preparation of the yeast extract from the yeast cultured by the yeast culture method of the present invention is not particularly limited, and any of the usual preparation methods may be used when preparing the yeast extract.
- the preparation method include a self-digestion method that solubilizes cells using a proteolytic enzyme or the like inherent in yeast cells, and an enzyme decomposition that solubilizes cells by adding an enzyme preparation derived from microorganisms or plants.
- hot water extraction method to solubilize cells by soaking in hot water for a certain period of time acid / alkaline decomposition method to solubilize cells by adding various acids or alkalis, freezing and thawing once
- acid / alkaline decomposition method to solubilize cells by adding various acids or alkalis
- freezing and thawing once
- There are a freeze-thaw method for crushing cells by performing the above a physical crushing method for crushing cells by physical stimulation, and the like.
- physical stimulation used in the physical crushing method include sonication, homogenization under high pressure, and grinding by mixing with a solid material such as glass beads.
- dry yeast cells having a high glutathione content can be obtained by subjecting the culture obtained by the yeast culture method of the present invention to a drying treatment.
- the method for drying the culture is not particularly limited, and any of the preparation methods commonly used for preparing dry yeast cells may be used. Examples of the preparation method include freeze-drying method, spray-drying method, and drum-drying method.
- a dry yeast powder having a high glutathione content and excellent handleability can be obtained.
- a fraction containing glutathione may be obtained from the culture obtained by the yeast culture method of the present invention.
- any method may be used as long as it is a commonly used method.
- an extract obtained by extraction with hot water or cell disruption is fractionated using an affinity column carrying a substance having a high affinity for sulfur-containing compounds, thereby containing glutathione at a high concentration. It becomes possible to concentrate and purify the fraction.
- the glutathione-rich yeast obtained by the yeast culture method of the present invention, the dried yeast cells of the yeast, the yeast extract prepared from the yeast, and the yeast extract powder may be used as a seasoning composition.
- the seasoning composition may consist only of the yeast extract of the present invention, and contains other components such as a stabilizer and a preservative in addition to the yeast extract of the present invention. It may be.
- the said seasoning composition can be suitably used for various food-drinks similarly to other seasoning compositions.
- the glutathione-rich yeast obtained by the yeast culture method of the present invention the dried yeast cells of the yeast, the yeast extract prepared from the yeast, and the yeast extract powder are directly contained in food and drink as raw materials. You can also. Thereby, the food / beverage products which contain glutathione in high concentration can be manufactured efficiently.
- These foods and drinks may be any foods and drinks that can normally be added with dry yeast, yeast extract, and seasoning compositions containing these, for example, alcoholic beverages, soft drinks, fermented foods, seasonings, soups. , Breads and confectionery.
- the said culture etc. can also be ingested as a supplement etc. by processing into a soft capsule, a hard capsule, and a tablet.
- the above yeast containing a high amount of glutathione, the dried yeast cell of the yeast, the yeast extract prepared from the yeast, and the yeast extract powder are added in the same manner as other raw materials.
- the food / beverage products which contain glutathione in high concentration can be manufactured.
- total glutathione content per dry cell weight (% (w / w))” may be rephrased as “GSH content (%)”.
- Saccharomyces cerevisiae YNN27 strain Saccharomyces cerevisiae BY4742 strain
- Saccharomyces cerevisiae NCYC506 strain Saccharomyces cerevisiae NCYC506 strain
- Schizosaccharomyces pombe wild strain JCM18 are strains JCM18. .
- Saccharomyces cerevisiae AB9 strain ⁇ MATa / ⁇ gpi10 / gpi10 ura3 / URA3 leu2 / LEU2> is a mutant strain having a mutation in the gene encoding ⁇ 1,2-mannosyltransferase and releasing mannan protein into the medium ( (For example, see the pamphlet of International Publication No.
- Saccharomyces cerevisiae AB9 strain has been deposited at the National Institute of Advanced Industrial Science and Technology Patent Biological Deposit Center (1-1-1 East Tsukuba, Ibaraki) (Accession number: FERM BP-10390) (date of deposit of the original deposit) : July 13, 2004, date of transfer to international deposit: August 3, 2005).
- Saccharomyces cerevisiae AB13 strain is a glutathione-rich mutant obtained as follows. First, a wild-type strain of Saccharomyces cerevisiae was subjected to EMS treatment, and mutant strains having a higher glutathione content than the parent strain were selected from the obtained mutant strains. Subsequently, the selected mutant strain was subjected to EMS treatment, and a mutant strain having a higher glutathione content than the parent strain was selected from the obtained mutant strains. By repeating this step twice or more, Saccharomyces cerevisiae AB13, KK101 strain, KK122 strain, and KK124 strain were obtained.
- compositions are shown in Tables 1 to 7, respectively.
- yeast was cultured by the yeast culturing method of the present invention using a semi-synthetic medium, and the GSH content (%) of the cultured yeast was measured.
- the KK101 strain stored in glycerol at ⁇ 80 ° C. was smeared on a YPD agar medium and cultured at 30 ° C. for 3 days. Thereafter, one loop of yeast grown on the YPD agar medium was inoculated into a 13-mL capacity tube containing 3 mL of YPD medium and cultured at 30 ° C. for 1 to 3 days at a stirring speed of 200 rpm.
- the obtained culture broth was used as a pre-culture broth and used for the following main culture.
- 150 ⁇ L of the preculture solution was inoculated into a 200 mL baffled Erlenmeyer flask containing 15 mL of a semi-synthetic medium supplemented with citric acid so as to be 0 to 100 mM, and cultured at 30 ° C. for 48 hours with a stirring speed of 200 rpm. did.
- the total amount of glutathione contained in the cells in the obtained culture is measured according to the method of Titze et al. (Analytical Biochemistry, Vol. 27, p502, 1969), and is divided by the dry cell weight to contain GSH.
- the rate (total glutathione content per dry cell weight) was calculated. Specifically, first, the obtained culture was centrifuged at 6000 rpm ⁇ 5 minutes at 4 ° C., and the recovered yeast cells were washed twice with purified water. After 2 mL of purified water is added and suspended in the yeast cells, 500 ⁇ L of this suspension is placed in an aluminum dish and dried at 105 ° C. for 5 hours or longer, and further left for 1 hour or longer in a desiccator, followed by drying.
- the weight was measured and used as the dry cell weight.
- 500 ⁇ L of the suspension was boiled in a hot water bath for 5 minutes, immediately cooled in an ice bath, centrifuged at 15000 rpm ⁇ 5 minutes at 4 ° C., and the recovered supernatant was used as a sample for glutathione determination. It was.
- a solution prepared by adding 5 ⁇ L of glutathione reductase and 500 ⁇ L of NADPH to 2.5 mL of 0.5 M potassium phosphate buffer (pH 7.0) mixed with 1 mM EDTA was placed in a 4 mL disposable cell immediately before the measurement.
- FIG. 1 is a graph showing the GSH content (%) for each citric acid concentration in the liquid medium. As a result, it was found that the GSH content (%) was increased by adding citric acid to the liquid medium. The GSH content (%) increased depending on the citric acid concentration. Under the conditions of this example, the maximum value was obtained at a citric acid concentration of 90 mM.
- yeast was cultured by the yeast culture method of the present invention using molasses medium (1), and the GSH content (%) of the cultured yeast was measured.
- Each conical flask was inoculated and cultured at 30 ° C. with shaking at 200 rpm for 48 hours.
- molasses medium originally contains several mM citric acid, the actual citric acid concentration in the molasses medium is slightly higher than the added citric acid concentration.
- the GSH content (%) of the cultured yeast was measured in the same manner as in Example 1. Furthermore, the pH of the liquid medium at the end of the culture was also measured. 2A and 2B show the citric acid in which the GSH content (%) of each culture (FIG. 2A) and the pH (final pH) of the liquid medium at the end of the culture (FIG. 2B) were added to the molasses medium (1). It is the graph shown for every density
- Example 3 When the Saccharomyces cerevisiae KK101 strain was cultured using a semi-synthetic medium to which various acids were added, the GSH content (%) of the yeast after the culture was measured, and each of the liquid medium with respect to the glutathione content of the yeast was measured. The effect of acid was investigated. 150 ⁇ L of the preculture solution prepared in the same manner as in Example 1 was inoculated into a 200 mL baffled Erlenmeyer flask containing 15 mL of a semi-synthetic medium to which various salts described in Table 8 were added so as to have the concentrations described in the table. The culture was performed at 30 ° C. with shaking at 200 rpm for 48 hours. The GSH content (%) of the cultured yeast was measured in the same manner as in Example 1. Furthermore, the pH (final pH) and OD 600 of the liquid medium at the end of the culture were also measured. The measurement results are shown in Tables 8 to 10.
- the pH of the liquid medium is adjusted to 4.0 to 6.0 by adding 50 mM or more of acid to the liquid medium, and in this addition amount range, It was found that the amount of added did not significantly affect the pH of the liquid medium. With phosphoric acid, it was found that the pH of the liquid medium increased depending on the amount added, and the effect of the amount added was large. In addition, when any liquid medium was used, the value of OD 600 at the end of the culture was about 30, and it was found that the effect of the type and amount of acid added to the liquid medium on the growth of yeast was small. .
- the GSH content was about 2.8% for acids other than citric acid, and there was no particular difference.
- GSH content rate was 2.4% and it was lower than others, but this is guessed because the pH of the liquid culture medium was less than 4.
- the GSH content tends to increase depending on the citric acid content. From these results, it is clear that the glutathione-rich effect obtained by culturing yeast in a citric acid-containing medium is an effect peculiar to citric acid.
- the semi-synthetic medium was used because the GSH content and the final pH of the liquid medium were almost the same when the citric acid concentration of the liquid medium was 50 mM and when other acids were added at 50 mM or more. In some cases, it is estimated that most of the citric acid added to have a citric acid concentration of 50 mM is used for adjusting the pH of the liquid medium.
- Example 4 For the Saccharomyces cerevisiae KK101 strain, yeast was cultured by the yeast culture method of the present invention using a liquid medium to which citric acid was further added with the pH adjusted, and the GSH content (% ) was measured. 150 ⁇ L of the preculture solution prepared in the same manner as in Example 1 was added to each 200 mL baffled Erlenmeyer flask containing 15 mL of semi-synthetic medium to which both acids were added so that the succinic acid concentration and citric acid concentration described in Table 11 were obtained. Inoculated and cultured at 30 ° C. with shaking at 200 rpm for 48 hours. The GSH content (%) of the cultured yeast was measured in the same manner as in Example 1.
- the pH (final pH) of the liquid medium at the end of the culture was also measured.
- Table 11 shows the measurement results.
- the citric acid was further added to the liquid medium, so that the GSH content was higher than when no addition was made. Increased dramatically.
- the GSH content which was 2.8% when the citric acid concentration was 0 mM (no addition)
- the citric acid concentration was 75 mM and 90 mM both were 4.3%, which was increased by 50% or more from the GSH content in the case of no addition.
- Example 3 Compared with the results of Example 3, even when the citric acid concentration in the liquid medium was the same, the present Example to which succinic acid was added together had a higher GSH content rate. It can be said that a part of the citric acid added to the medium was used to control the pH of the liquid medium. That is, from these results, by adding citric acid to the liquid medium whose pH was controlled to 4.0 to 6.0, citric acid was added to the liquid medium whose pH was not controlled to 4.0 to 6.0. It is apparent that a higher glutathione-rich effect can be obtained with a smaller amount of citric acid added than when adding.
- Saccharomyces cerevisiae KK122 and KK124 strains were cultured with the yeast culture method of the present invention under the culture conditions adjusted for pH using a jar fermenter, and the GSH content (% ) was measured.
- pre-cultures of Saccharomyces cerevisiae KK122 strain and KK124 strain were prepared. 100 ⁇ L of each pre-culture solution was inoculated into a 200 mL baffled Erlenmeyer flask containing 10 mL of YPD medium, and cultured at 30 ° C. for 1 day at a stirring speed of 200 rpm. The obtained culture broth was used as a pre-culture broth and used for the following main culture.
- 3A and 3B are diagrams showing the measurement results of GSH content and OD 600 of Saccharomyces cerevisiae KK122 strain [FIG. 3A] and KK124 strain [FIG. 3B].
- the bar graph is the result of GSH content (%)
- the line graph is the result of OD 600 .
- the results of “90 mM citric acid” cultured in a 90 mM citric acid-containing medium and the results of “0 mM citric acid (no addition)” cultured in a semi-synthetic medium are shown.
- Saccharomyces cerevisiae KK122 strain and KK124 strain also had a high GSH content by culturing in a liquid medium containing citric acid, like the Saccharomyces cerevisiae KK101 strain.
- the slope of the OD 600 graph was slightly smaller than when cultured in a semi-synthetic medium, but the OD 600 values at the end of the culture were similar. Met. From this, it can be observed that by adding citric acid to the liquid medium, although a slight delay in the rise is observed, the yeast can eventually be cultured in the same manner as in the case of no addition of citric acid. It was confirmed that the effect on growth was small.
- Example 6 For various Saccharomyces spp., Yeast was cultured by the yeast culture method of the present invention using a semi-synthetic medium, and the GSH content (%) of the cultured yeast was measured. Based on the results of Example 3, when the citric acid concentration was 50 mM as a reference (control), and an increase in the GSH content rate was observed depending on the citric acid concentration, the effect of high glutathione content of the present invention was It was confirmed that it was played. First, by the same method as in Example 1, precultures of the strains listed in Table 12 were prepared.
- Example 7 For various Saccharomyces spp., Yeast was cultured by the yeast culture method of the present invention using a YPD medium, and the GSH content (%) of the cultured yeast was measured. When an increase in the GSH content rate was observed depending on the citric acid concentration, it was confirmed that the effect of high glutathione content of the present invention could be confirmed. First, by the same method as in Example 1, precultures of the strains listed in Table 13 were prepared.
- each preculture was inoculated into a 200 mL baffled Erlenmeyer flask containing 15 mL of YPD medium supplemented appropriately with citric acid so that the citric acid concentration was 0 to 90 mM, and stirred at 30 ° C.
- the culture was shaken at 200 rpm for 48 hours.
- the GSH content (%) of the cultured yeast was measured in the same manner as in Example 1. Table 13 shows the measurement results. As a result, it was confirmed that each strain of Saccharomyces cerevisiae has the glutathione high content effect of the present invention in both the mutant strain and the wild strain in the YPD medium.
- Yeasts other than Saccharomyces were also cultured by the yeast culturing method of the present invention, and the GSH content (%) of the cultured yeast was measured.
- Candida utilis 1561 strain was cultured in a YPD medium containing citric acid, and the GSH content rate was measured.
- a preculture solution of Candida utilis 1561 strain was prepared by the same method as in Example 1.
- 15 ⁇ L of each preculture was inoculated into a 200 mL baffled Erlenmeyer flask containing 15 mL of YPD medium appropriately added with citric acid so that the citric acid concentration was 0 to 62.5 mM (the inoculation rate was 0. 0).
- Example 14 shows the measurement results. As a result, it was confirmed that the Candida utilis 1561 strain also has the glutathione high content effect of the present invention as in the case of the genus Saccharomyces.
- Yeast was also cultured with respect to yeast of other strains belonging to the genus Candida by the yeast culture method of the present invention, and the GSH content (%) of the cultured yeast was measured.
- Candida utilis 1560 strain was cultured in a molasses medium (2) containing citric acid, and the GSH content rate was measured.
- a preculture solution of Candida utilis 1560 strain was prepared by the same method as in Example 1.
- 45 ⁇ L of each preculture was inoculated into a 200 mL baffled Erlenmeyer flask containing 15 mL of molasses medium (2) with appropriate addition of citric acid so that the citric acid concentration was 0-62.5 mM (inoculation magnification).
- Example 15 shows the measurement results. As a result, it was confirmed that the Candida utilis 1560 strain also has the glutathione high content effect of the present invention as in the case of the genus Saccharomyces.
- Example 10 For Saccharomyces spp., Yeast was cultured by the yeast culture method of the present invention using an SD medium, and the GSH content (%) of the cultured yeast was measured. When an increase in the GSH content rate was observed depending on the citric acid concentration, it was confirmed that the effect of high glutathione content of the present invention could be confirmed. First, by the same method as in Example 1, precultures of the strains described in Table 16 were prepared.
- each preculture was inoculated into a 200 mL baffled Erlenmeyer flask containing 15 mL of YPD medium supplemented appropriately with citric acid so that the citric acid concentration was 50 to 150 mM, and stirred at 30 ° C.
- the culture was shaken at 200 rpm for 48 hours.
- the GSH content (%) of the cultured yeast was measured in the same manner as in Example 1. The measurement results are shown in Table 16.
- each strain of Saccharomyces cerevisiae has the glutathione-rich effect of the present invention in both the mutant and wild strains even in the SD medium. From these results, the glutathione high content effect of the present invention is not particularly limited in the composition of the liquid medium to be used, and any medium can be used by adding a sufficient amount of citric acid. It can be said.
- Example 11 After the cultivation was started, the citrate concentration of the liquid medium was adjusted to 20 mM or more, whereby yeast was cultured by the yeast cultivation method of the present invention, and the GSH content (%) of the cultured yeast was measured. Based on the results of Example 3, when the citric acid concentration at the start of culture is 50 mM as a reference (control), when the GSH content is higher than this, the effect of high glutathione content of the present invention is achieved. It can be confirmed that.
- precultures of Saccharomyces cerevisiae KK101 strain, KK122 strain, KK124 strain, and AB13 strain were prepared. Subsequently, each strain was cultured as follows.
- citric acid was added to one of the remaining two to which citric acid was not added so that the citric acid concentration was 90 mM, and the culture was continued as it was (after 6 hours). 90 mM sample added).
- citric acid was added so that the citric acid concentration was 90 mM to the remaining one not added with citric acid (90 mM sample added after 9 hours), and the culture was continued as it was.
- the culture was completed in all the flasks to obtain 5 samples. The same culture operation was performed on each strain to obtain a total of 20 samples.
- FIG. 4 is a diagram showing the results of measuring the GSH content (%) of yeast in each sample.
- the GSH content (%) was measured in the same manner as in Example 1.
- the GSH content rate of the sample to which 90 mM citric acid was added was higher than that of the 50 mM sample added at the initial stage, regardless of the addition time of citric acid. That is, from these results, even when the concentration of citric acid in the liquid medium is adjusted to a sufficient concentration to achieve the glutathione-rich effect of the present invention after the start of culture, the glutathione content of the yeast is reduced. It was confirmed that it could be increased.
- the initial addition 90 mM sample has the highest GSH content rate, and the later the addition time of citric acid, the lower the GSH content rate. Since the tendency was observed, it was considered that it is preferable to adjust the citric acid concentration after the start of the culture at least within 9 hours after the start of the culture, that is, at a time when budding of the yeast is active.
- Example 1 It was examined whether or not citric acid added to the liquid medium was assimilated as a carbon source.
- a preculture solution of Saccharomyces cerevisiae KK101 strain was prepared by the same method as in Example 1.
- a semi-synthetic medium 50 mM citric acid-containing medium
- semi-synthetic added with sodium citrate so that the citric acid concentration is 90 mM.
- FIG. 5 is a diagram showing measurement results of citric acid content in each supernatant before and after culture.
- 50 mM Na Citrate is the result of culturing in a medium containing 50 mM citric acid
- 90 mM Na Citrate is the result of culturing in a medium containing 90 mM citric acid
- 100 mM K Phospate is cultured in a medium containing 100 mM phosphoric acid. The results are shown respectively.
- the yeast culturing method of the present invention can easily increase the glutathione content of yeast such as Saccharomyces, so that the culturing method can be used particularly in the food field.
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Abstract
Description
本願は2010年3月26日に日本に出願された特願2010-073818号に基づき優先権を主張し、その内容をここに援用する。
特許文献1に記載の方法のように、変異処理等を行い遺伝的な改変が施された酵母の中から、よりグルタチオン含有量の高い酵母をスクリーニングする方法によっても、グルタチオン高含有酵母を得ることができるが、変異処理及びスクリーニングは手間と労力を要し、また、必ずしもグルタチオン高含有酵母を得ることができない場合も多い。
また、組換え体よりも天然の酵母(野生株)が求められる場合があり、変異処理を行うことなく、酵母中のグルタチオン含有量を高める方法の開発が望まれている。例えば、酵母自体や酵母エキスを食用として用いる場合には、組換え体よりも野生株が好まれる場合がある。
(1) 酵母を、クエン酸濃度が20mM以上である液体培地中で培養することを含む、酵母の培養方法、
(2) 前記液体培地のクエン酸濃度が200mM以下である前記(1)記載の酵母の培養方法、
(3) 培養開始時における液体培地のクエン酸濃度が20mM以上である前記(1)又は(2)記載の酵母の培養方法、
(4) 培養開始時における液体培地のクエン酸濃度が20mM未満であり、かつ、酵母の増殖状態が誘導期又は対数増殖期に、液体培地のクエン酸濃度を20~200mMに調整することを含む、前記(1)記載の酵母の培養方法、
(5) 培養開始時における液体培地のクエン酸濃度が20mM未満であり、かつ、培養開始後9時間以内に、液体培地のクエン酸濃度を20~200mMに調整することを含む、前記(1)記載の酵母の培養方法。
(6) 前記酵母がサッカロマイセス(Saccharomyces)属菌又はキャンディタ(Candida)属菌である前記(1)~(5)のいずれか1項記載の酵母の培養方法、
(7) 前記酵母が、サッカロマイセス・セレビシエ(Saccharomyces cerevisiae)又はキャンディダ・ユティリス(Candida utilis)である前記(1)~(5)のいずれか1項記載の酵母の培養方法、
(8) 酵母を、クエン酸濃度が20mM以上である液体培地中で培養することを含む、酵母のグルタチオン含有量を高める方法、
(9) 前記(1)~(7)のいずれか1項記載の酵母の培養方法で培養された酵母を回収することを含む、酵母の製造方法、
(10) 前記(1)~(7)のいずれか1項記載の酵母の培養方法で培養された酵母から、酵母エキスを抽出することを含む、酵母エキスの製造方法、
(11) 前記(1)~(7)のいずれか1項記載の酵母の培養方法で培養された酵母、及び前記酵母から調製された酵母エキスからなる群より選択される1以上の製造物を原料として用いることを含む、飲食品の製造方法、
(12) 前記(1)~(7)のいずれか1項記載の酵母の培養方法で培養された酵母から調製される、酵母エキス、
(13) 前記(1)~(7)のいずれか1項記載の酵母の培養方法で培養された酵母、又は前記(12)記載の酵母エキスを含有する、調味料組成物、
(14) 前記(1)~(7)のいずれか1項記載の酵母の培養方法で培養された酵母、又は前記(12)記載の酵母エキスを含有する、飲食品、
を提供するものである。
本発明のグルタチオン高含有酵母とは、親株と比較して菌体内のグルタチオン含量が優位に増加している酵母をいう。
また培養物とは、酵母菌体及び酵母の培養に用いた培地を含む培養物を意味する。
液体培地のクエン酸濃度を調整する方法においては、液体培地に固体のクエン酸を添加し調整してもよく、クエン酸水溶液を添加し調整してもよい。
なお、対数増殖期は、回分及び流加培養においては、吸光度等を指標として培養容器中の酵母の量を経時的に測定した場合に、対数的に増大することが観察される時期である。
一方、連続培養においては、培養容器中の酵母の量がほぼ一定であることが観察される時期である。
誘導期は、回分及び流加培養において、培養開始後から対数増殖期に至る前の時期である。
一方、連続培養においては、メルクマールとなるパラメータを一定値に制御するまでの時期である。
例えば、培養開始から9時間以内、好ましくは3時間以内に液体培地のクエン酸濃度を調整することにより、出芽が盛んな状態の酵母に対して十分な濃度のクエン酸を作用させることができる。
クエン酸が添加される液体培地としては、炭素源、窒素源、及び無機塩等を含んでいるものであって、通常サッカロマイセス・セレビシエ等の酵母の培養に用いられるいずれの培地も用いることができる。
前記培養物から酵母を回収する方法は、特に限定されるものではなく、酵母の培養物から酵母を回収する場合に通常行なわれている回収方法のいずれを用いてもよい。前記培養物から酵母を回収する方法として、例えば、酵母の培養物を遠心分離処理する方法等が挙げられる。
前記調味料組成物は、他の調味料組成物と同様に、様々な飲食品に適宜用いることができる。
サッカロマイセス・セレビシエKK101株に対して、半合成培地を用いて本発明の酵母の培養方法により酵母の培養を行ない、培養後の酵母のGSH含有率(%)を測定した。
まず、-80℃でグリセロール中に保存していたKK101株を、YPD寒天培地上に塗沫して30℃にて3日間培養した。その後、YPD寒天培地に生育した酵母1ループ分を、3mLのYPD培地を含む13mL容アシストチューブに接種し、30℃にて、攪拌速度200rpmで1~3日間、振盪培養した。得られた培養液を、前培養液とし、以下の本培養に用いた。
150μLの前培養液を、0~100mMとなるようにクエン酸を添加した半合成培地を15mL含む200mL容バッフル付三角フラスコにそれぞれ接種し、30℃にて、攪拌速度200rpmで48時間、振盪培養した。
サッカロマイセス・セレビシエAB13株に対して、糖蜜培地(1)を用いて本発明の酵母の培養方法により酵母の培養を行ない、培養後の酵母のGSH含有率(%)を測定した。
実施例1と同様にして調製した150μLの前培養液(1)を、添加されたクエン酸の濃度が0~60mMとなるようにクエン酸を添加した糖蜜培地(1)を15mL含む200mL容バッフル付三角フラスコにそれぞれ接種し、30℃にて、攪拌速度200rpmで48時間、振盪培養した。糖蜜培地中には元々数mMのクエン酸が含まれているため、実際の糖蜜培地中のクエン酸濃度は、添加されたクエン酸濃度よりも若干高くなる。培養後の酵母のGSH含有率(%)を、実施例1と同様にして測定した。さらに、培養終了時点における液体培地のpHも測定した。
図2A及び図2Bは、各培養物のGSH含有率(%)〔図2A〕及び培養終了時点における液体培地のpH(終pH)〔図2B〕を、糖蜜培地(1)に添加したクエン酸濃度ごとに示したグラフである。この結果、糖蜜培地(1)に対しても、クエン酸を10mM以上添加することにより、酵母のGSH含有率(%)を、添加前よりも10%以上増大させることができた。また、クエン酸を添加することにより、培養終了時点の液体培地のpHが上昇していたが、添加するクエン酸量が20mM以上の場合には、pH5.2前後で安定していた。
サッカロマイセス・セレビシエKK101株を、様々な酸を添加した半合成培地を用いて培養した場合の、培養後の酵母のGSH含有率(%)を測定し、酵母のグルタチオン含有量に対する液体培地中の各酸の影響を調べた。
実施例1と同様にして調製した150μLの前培養液を、表8記載の各種の塩を表記載の濃度となるように添加した半合成培地を15mL含む200mL容バッフル付三角フラスコにそれぞれ接種し、30℃にて、攪拌速度200rpmで48時間、振盪培養した。
培養後の酵母のGSH含有率(%)を、実施例1と同様にして測定した。さらに、培養終了時点における液体培地のpH(終pH)及びOD600も測定した。測定結果を表8~10に示す。
また、いずれの液体培地を用いた場合も、培養終了時のOD600の値は30前後であり、酵母の生育性に対する、液体培地に添加する酸の種類や量の影響は小さいことが分かった。
これらの結果から、クエン酸含有培地で酵母を培養することにより得られるグルタチオン高含有効果は、クエン酸に特有の効果であることが明らかである。なお、液体培地のクエン酸濃度を50mMとした場合と、他の酸を50mM以上添加した場合とで、GSH含有率と液体培地の終pHがほぼ同等であったことから、半合成培地を用いる場合には、クエン酸濃度が50mMとなるように添加されたクエン酸のうちの大部分は、液体培地のpHの調整に利用されていると推定される。
サッカロマイセス・セレビシエKK101株に対して、pHを調整した状態でさらにクエン酸を添加した液体培地を用いて本発明の酵母の培養方法により酵母の培養を行ない、培養後の酵母のGSH含有率(%)を測定した。
実施例1と同様にして調製した150μLの前培養液を、表11記載のコハク酸濃度及びクエン酸濃度となるように両酸を添加した半合成培地を15mL含む200mL容バッフル付三角フラスコにそれぞれ接種し、30℃にて、攪拌速度200rpmで48時間、振盪培養した。培養後の酵母のGSH含有率(%)を、実施例1と同様にして測定した。
さらに、培養終了時点における液体培地のpH(終pH)も測定した。測定結果を表11に示す。
この結果、コハク酸を添加することによって液体培地のpHを5.5~6.0に調整した状態で、液体培地にさらにクエン酸を添加することにより、無添加の場合よりもGSH含有率が飛躍的に増大した。具体的には、クエン酸濃度が0mM(無添加)の場合は2.8%であったGSH含有率が、クエン酸濃度が50mMの場合には3.7%となり、30%以上増大した。一方、クエン酸濃度が75mM及び90mMの場合にはいずれも4.3%となり、無添加の場合のGSH含有率よりも50%以上増大した。
実施例3の結果と比較すると、液体培地中のクエン酸濃度が同じ場合でも、コハク酸をともに添加した本実施例のほうが、GSH含有率が高く、この比較からも、実施例3では、液体培地に添加されたクエン酸の一部が液体培地のpHを制御することに利用されていたといえる。
すなわち、これらの結果から、pHが4.0~6.0に制御された液体培地にクエン酸を添加することにより、pHが4.0~6.0に制御されていない液体培地にクエン酸を添加した場合よりも、より少ないクエン酸添加量でより高いグルタチオン高含有効果が得られることが明らかである。
サッカロマイセス・セレビシエKK122株及びKK124株に対して、ジャーファーメンターを用いてpHを調整した培養条件で、本発明の酵母の培養方法により酵母の培養を行ない、培養後の酵母のGSH含有率(%)を測定した。
まず、実施例1と同様の方法により、サッカロマイセス・セレビシエKK122株及びKK124株の前々培養液を、それぞれ調製した。各前々培養液100μLを、10mLのYPD培地を含む200mL容バッフル付三角フラスコに接種し、30℃にて、攪拌速度200rpmで1日間、振盪培養した。得られた培養液を、前培養液とし、以下の本培養に用いた。
10mLの各前培養液を、半合成培地又はクエン酸濃度が90mMとなるようにクエン酸ナトリウムを添加した半合成培地(90mMクエン酸含有培地)を1L含む2L容ジャーファーメンターに接種し、初発pHが6.8、攪拌速度300rpm、通気量1V.V.M.、30℃の条件で、48時間、通気攪拌培養した。培養中、pHの下限値が6.0となるように、1N 水酸化ナトリウム溶液で調整した。培養開始から12時間ごとにサンプリングし、OD600を測定した。また、培養開始後24、36、及び48時間にサンプリングしたサンプルについては、実施例1と同様にしてGSH含有率も測定した。
図3A及び3Bは、サッカロマイセス・セレビシエKK122株〔図3A〕及びKK124株〔図3B〕の、GSH含有率及びOD600の測定結果を示した図である。図中、棒グラフがGSH含有率(%)の結果であり、折れ線グラフがOD600の結果である。また、「90mM クエン酸」が90mMクエン酸含有培地で培養した結果を、「0mM クエン酸(無添加)」が半合成培地で培養した結果を、それぞれ示す。この結果、サッカロマイセス・セレビシエKK122株及びKK124株についても、サッカロマイセス・セレビシエKK101株と同様に、クエン酸を含有する液体培地中で培養することにより、GSH含有率が高くなった。また、両株とも、90mMクエン酸含有培地で培養した場合には、半合成培地で培養した場合よりも、若干OD600グラフの傾きが小さかったが、培養終了時点のOD600値は、同程度であった。このことから、液体培地にクエン酸を添加することにより、若干立ち上がりに遅れが観察されるものの、最終的にはクエン酸無添加の場合と同じように酵母は培養でき、クエン酸添加による酵母の生育性への影響は小さいといえることが確認された。
様々なサッカロマイセス属菌に対して、半合成培地を用いて本発明の酵母の培養方法により酵母の培養を行ない、培養後の酵母のGSH含有率(%)を測定した。実施例3の結果に基づき、クエン酸濃度を50mMとした場合を基準(対照)とし、クエン酸濃度依存的にGSH含有率の増大が観察された場合には、本発明のグルタチオン高含有効果が奏されていることが確認できるとした。
まず、実施例1と同様の方法により、表12に記載の菌株の前培養液をそれぞれ調製した。次いで、150μLの各前培養液を、50~150mMとなるようにクエン酸を添加した半合成培地を15mL含む200mL容バッフル付三角フラスコにそれぞれ接種し、30℃にて、攪拌速度200rpmで48時間、振盪培養した。培養後の酵母のGSH含有率(%)を、実施例1と同様にして測定した。
測定結果を表12に示す。この結果、変異株であるサッカロマイセス・セレビシエKK101株、KK122株、KK124株、及びAB13株のみならず、野生株であるサッカロマイセス・セレビシエYNN27株、BY4742株、及びNCYC506株のいずれにおいても、クエン酸濃度が75~150mMの場合に、50mMの場合よりもGSH含有率が10%以上高くなっていた。これらの結果から、本発明の酵母の培養方法は、特定の変異株のみならず、野生株に対して適用した場合でも、本発明のグルタチオン高含有効果が得られることが明らかである。
様々なサッカロマイセス属菌に対して、YPD培地を用いて本発明の酵母の培養方法により酵母の培養を行ない、培養後の酵母のGSH含有率(%)を測定した。クエン酸濃度依存的にGSH含有率の増大が観察された場合には、本発明のグルタチオン高含有効果が奏されていることが確認できるとした。
まず、実施例1と同様の方法により、表13に記載の菌株の前培養液をそれぞれ調製した。次いで、150μLの各前培養液を、クエン酸濃度が0~90mMとなるように適宜クエン酸を添加したYPD培地を15mL含む200mL容バッフル付三角フラスコにそれぞれ接種し、30℃にて、攪拌速度200rpmで48時間、振盪培養した。培養後の酵母のGSH含有率(%)を、実施例1と同様にして測定した。
測定結果を表13に示す。この結果、サッカロマイセス・セレビシエの各菌株は、変異株と野生株のいずれも、YPD培地においても本発明のグルタチオン高含有効果が得られることが確認された。
サッカロマイセス属以外の酵母に対しても本発明の酵母の培養方法により酵母の培養を行ない、培養後の酵母のGSH含有率(%)を測定した。具体的には、キャンディダ・ユティリス1561株を、クエン酸を含有させたYPD培地で培養し、GSH含有率を測定した。
まず、実施例1と同様の方法により、キャンディダ・ユティリス1561株の前培養液を調製した。次いで、15μLの各前培養液を、クエン酸濃度が0~62.5mMとなるように適宜クエン酸を添加したYPD培地を15mL含む200mL容バッフル付三角フラスコにそれぞれ接種し(接種倍率が0.1%)、30℃にて、攪拌速度200rpmで40時間、振盪培養した。培養後の酵母のGSH含有率(%)を、実施例1と同様にして測定した。
測定結果を表14に示す。この結果、キャンディダ・ユティリス1561株においても、サッカロマイセス属菌と同様に本発明のグルタチオン高含有効果が得られることが確認された。
キャンディダ属に属する他の菌株の酵母に対しても本発明の酵母の培養方法により酵母の培養を行ない、培養後の酵母のGSH含有率(%)を測定した。具体的には、キャンディダ・ユティリス1560株を、クエン酸を含有させた糖蜜培地(2)で培養し、GSH含有率を測定した。
まず、実施例1と同様の方法により、キャンディダ・ユティリス1560株の前培養液を調製した。次いで、45μLの各前培養液を、クエン酸濃度が0~62.5mMとなるように適宜クエン酸を添加した糖蜜培地(2)を15mL含む200mL容バッフル付三角フラスコにそれぞれ接種し(接種倍率が0.3%)、30℃にて、攪拌速度200rpmで24時間、振盪培養した。培養後の酵母のGSH含有率(%)を、実施例1と同様にして測定した。
測定結果を表15に示す。この結果、キャンディダ・ユティリス1560株においても、サッカロマイセス属菌と同様に本発明のグルタチオン高含有効果が得られることが確認された。
サッカロマイセス属菌に対して、SD培地を用いて本発明の酵母の培養方法により酵母の培養を行ない、培養後の酵母のGSH含有率(%)を測定した。クエン酸濃度依存的にGSH含有率の増大が観察された場合には、本発明のグルタチオン高含有効果が奏されていることが確認できるとした。
まず、実施例1と同様の方法により、表16に記載の菌株の前培養液をそれぞれ調製した。次いで、150μLの各前培養液を、クエン酸濃度が50~150mMとなるように適宜クエン酸を添加したYPD培地を15mL含む200mL容バッフル付三角フラスコにそれぞれ接種し、30℃にて、攪拌速度200rpmで48時間、振盪培養した。培養後の酵母のGSH含有率(%)を、実施例1と同様にして測定した。
測定結果を表16に示す。この結果、サッカロマイセス・セレビシエの各菌株は、変異株と野生株のいずれも、SD培地においても本発明のグルタチオン高含有効果が得られることが確認された。これらの結果から、本発明のグルタチオン高含有効果は、用いる液体培地の組成は特に限定されるものではなく、十分量のクエン酸を添加することにより、いずれの培地を用いても効果が得られるといえる。
培養開始後に、液体培地のクエン酸濃度を20mM以上に調整することにより、本発明の酵母の培養方法により酵母の培養を行ない、培養後の酵母のGSH含有率(%)を測定した。実施例3の結果に基づき、培養開始時点におけるクエン酸濃度を50mMとした場合を基準(対照)とし、これよりもGSH含有率が高い場合には、本発明のグルタチオン高含有効果が奏されていることが確認できるとした。
実施例1と同様の方法により、サッカロマイセス・セレビシエKK101株、KK122株、KK124株、及びAB13株の前培養液を、それぞれ調製した。
次いで、各菌株に対して、それぞれ次のように培養を行った。まず、150μLの前培養液を、半合成培地を15mL含む200mL容バッフル付三角フラスコ5本にそれぞれ接種した。このうちの2本のフラスコには、それぞれ、クエン酸濃度が50mM又は90mMとなるように、それぞれクエン酸を添加した(初期添加50mMサンプル、及び初期添加90mMサンプル)。これらの5本のフラスコを、30℃にて、攪拌速度200rpmで振盪培養した。培養開始から3時間後、クエン酸を添加していない3本のうちの1本に、クエン酸濃度が90mMとなるようにクエン酸を添加し、そのまま培養を続けた(3時間後添加90mMサンプル)。さらに、培養開始から6時間後に、クエン酸を添加していない残る2本のうちの1本に、クエン酸濃度が90mMとなるようにクエン酸を添加し、そのまま培養を続けた(6時間後添加90mMサンプル)。最後に、培養開始から9時間後に、クエン酸を添加していない残る1本に、クエン酸濃度が90mMとなるようにクエン酸を添加し(9時間後添加90mMサンプル)、そのまま培養を続け、培養開始から48時間後に全てのフラスコで培養を終了し、5サンプルを得た。各菌株に対して同様の培養操作を行い、合計20サンプルを得た。
なお、GSH含有率(%)の測定は、実施例1と同様にして行った。この結果、全ての菌株において、クエン酸の添加時期に関わらず、90mMのクエン酸を添加したサンプルのGSH含有率は、初期添加50mMサンプルよりも高いことが確認された。つまり、これらの結果から、培養開始後に、液体培地のクエン酸濃度を、本発明のグルタチオン高含有効果が奏されるために十分な濃度に調整した場合であっても、酵母のグルタチオン含有量を高められることが確認された。また、菌株ごとに多少の差異はあるが、基本的にいずれの菌株においても、初期添加90mMサンプルが最もGSH含有率が高く、かつ、クエン酸の添加時期が遅くなるほど、GSH含有率が低くなる傾向が観察されたことから、培養開始後にクエン酸濃度を調整する場合には、少なくとも培養開始後9時間以内、つまり酵母の出芽が盛んな時期に行うことが好ましいと考えられた。
液体培地に添加されたクエン酸が、炭素源として資化されているかどうかを調べた。
まず、実施例1と同様の方法により、サッカロマイセス・セレビシエKK101株の前培養液を調製した。
150μLの前培養液を、クエン酸濃度が50mMとなるようにクエン酸ナトリウムを添加した半合成培地(50mMクエン酸含有培地)、クエン酸濃度が90mMとなるようにクエン酸ナトリウムを添加した半合成培地(90mMクエン酸含有培地)、又はリン酸濃度が100mMとなるようにリン酸カリウムを添加した半合成培地(100mMりン酸含有培地)を15mL含む200mL容バッフル付三角フラスコに接種し、30℃にて、攪拌速度200rpmで48時間、振盪培養した。酵母接種後培養前に培養上清の一部を予め採取しておき、培養終了後の上清とともに、これらの上清中のクエン酸の含有量を測定した。クエン酸量の測定は、市販のキット(商品名:F-キットクエン酸、Roche社製)を用いたF-kit法により行った。
図5は、培養前後の各上清中のクエン酸含有量の測定結果を示した図である。図中、「50mM Na Citrate」は50mMクエン酸含有培地で培養した結果を、「90mM Na Citrate」は90mMクエン酸含有培地で培養した結果を、「100mM K Phospate」は100mMリン酸含有培地で培養した結果を、それぞれ示す。この結果、いずれの液体培地を用いた場合でも、上清中のクエン酸含有量は、培養前よりも培養後において若干増大していた。これは、培養工程において、酵母がクエン酸を生成したためと考えられる。つまり、これらの結果から、酵母は、液体培地中のクエン酸を資化しておらず、液体培地に添加されたクエン酸が炭素源として用いられていないことが明らかである。
Claims (14)
- 酵母を、クエン酸濃度が20mM以上である液体培地中で培養することを含む、酵母の培養方法。
- 前記液体培地のクエン酸濃度が200mM以下である請求項1記載の酵母の培養方法。
- 培養開始時における液体培地のクエン酸濃度が20mM以上である請求項1又は2記載の酵母の培養方法。
- 培養開始時における液体培地のクエン酸濃度が20mM未満であり、かつ、
酵母の増殖状態が誘導期又は対数増殖期に、液体培地のクエン酸濃度を20~200mMに調整することを含む、請求項1記載の酵母の培養方法。 - 培養開始時における液体培地のクエン酸濃度が20mM未満であり、かつ、
培養開始後9時間以内に、液体培地のクエン酸濃度を20~200mMに調整することを含む、請求項1記載の酵母の培養方法。 - 前記酵母がサッカロマイセス(Saccharomyces)属菌又はキャンディタ(Candida)属菌である請求項1~5のいずれか1項記載の酵母の培養方法。
- 前記酵母が、サッカロマイセス・セレビシエ(Saccharomyces cerevisiae)又はキャンディダ・ユティリス(Candida utilis)である請求項1~5のいずれか1項記載の酵母の培養方法。
- 酵母を、クエン酸濃度が20mM以上である液体培地中で培養することを含む、酵母のグルタチオン含有量を高める方法。
- 請求項1~7のいずれか1項記載の酵母の培養方法で培養された酵母を回収することを含む、酵母の製造方法。
- 請求項1~7のいずれか1項記載の酵母の培養方法で培養された酵母から、酵母エキスを抽出することを含む、酵母エキスの製造方法。
- 請求項1~7のいずれか1項記載の酵母の培養方法で培養された酵母、及び前記酵母から調製された酵母エキスからなる群より選択される1以上の製造物を原料として用いることを含む、飲食品の製造方法。
- 請求項1~7のいずれか1項記載の酵母の培養方法で培養された酵母から調製される酵母エキス。
- 請求項1~7のいずれか1項記載の酵母の培養方法で培養された酵母、又は請求項12記載の酵母エキスを含有する調味料組成物。
- 請求項1~7のいずれか1項記載の酵母の培養方法で培養された酵母、又は請求項12記載の酵母エキスを含有する飲食品。
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CN102776258A (zh) * | 2012-08-13 | 2012-11-14 | 苏州大学 | 一种s-腺苷-l-蛋氨酸和谷胱甘肽联产发酵方法 |
WO2013146465A1 (ja) * | 2012-03-30 | 2013-10-03 | オリエンタル酵母工業株式会社 | 銅高含有酵母抽出物及びその製造方法、並びに、食品、及び野菜の緑色保持復元剤 |
WO2015151867A1 (ja) * | 2014-03-31 | 2015-10-08 | 興人ライフサイエンス株式会社 | グルタチオンを含む酵母抽出物のメラニン産生抑制剤としての利用 |
JP2019137655A (ja) * | 2018-02-14 | 2019-08-22 | 共栄化学工業株式会社 | 皮膚外用剤 |
WO2020050273A1 (ja) * | 2018-09-07 | 2020-03-12 | 株式会社カネカ | グルタチオンの製造方法 |
CN114395493A (zh) * | 2022-01-17 | 2022-04-26 | 广东中烟工业有限责任公司 | 一种改善上部烟叶品质的发酵液及其应用 |
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Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS59151894A (ja) | 1983-02-18 | 1984-08-30 | Kohjin Co Ltd | グルタチオン高含有酵母の製造法 |
JP2006042638A (ja) * | 2004-08-02 | 2006-02-16 | Asahi Breweries Ltd | 酵母変異株、グルタチオン高含有酵母の製造方法、その培養物、その分画物、酵母エキスおよびグルタチオン含有飲食品 |
WO2006025295A1 (ja) | 2004-08-30 | 2006-03-09 | Asahi Breweries, Ltd. | マンナンタンパク質を放出する酵母株およびマンナンタンパク質の製造法 |
JP2010073818A (ja) | 2008-09-17 | 2010-04-02 | Canon Inc | 測定装置、露光装置およびデバイス製造方法 |
Family Cites Families (10)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS5191393A (en) * | 1975-12-25 | 1976-08-10 | Hatsukohonyoru ll hisuchijinnoseizoho | |
JPS60196185A (ja) * | 1984-03-19 | 1985-10-04 | Chemo Sero Therapeut Res Inst | 形質転換酵母の培養方法 |
JP2816422B2 (ja) * | 1992-11-08 | 1998-10-27 | 株式会社コスモ総合研究所 | 微生物による5−アミノレブリン酸の製造方法 |
GB9404270D0 (en) * | 1994-03-05 | 1994-04-20 | Delta Biotechnology Ltd | Yeast strains and modified albumins |
JP2000279164A (ja) * | 1999-03-31 | 2000-10-10 | Kohjin Co Ltd | 酵母の製造方法 |
MY128920A (en) * | 2000-05-25 | 2007-02-28 | Ajinomoto Kk | METHOD FOR PRODUCING y-GLUTAMYLCYSTEINE |
BRPI0513617A (pt) * | 2004-08-02 | 2008-05-13 | Asahi Breweries Ltd | grupo de levedura mutante, método para produção de levedura rica em glutationa, cultura e fração da mesma, extrato de levedura, células de levedura seca e alimento e bebida contendo glutationa |
JP4412658B2 (ja) * | 2004-08-31 | 2010-02-10 | 国立大学法人京都大学 | チオレドキシン高含有酵母およびその製造法 |
CN101121922B (zh) * | 2006-08-08 | 2010-11-10 | 中国人民解放军第三○二医院 | 酿酒酵母细胞培养分泌液制备方法及其抗dna病毒和促肝细胞生长的用途 |
JP2009107962A (ja) * | 2007-10-29 | 2009-05-21 | Asahi Breweries Ltd | 酵母エキス含有組成物 |
-
2011
- 2011-03-25 CN CN201180013517.9A patent/CN102812119B/zh active Active
- 2011-03-25 DK DK11759611.4T patent/DK2554656T3/en active
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- 2011-03-25 EP EP11759611.4A patent/EP2554656B1/en active Active
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Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS59151894A (ja) | 1983-02-18 | 1984-08-30 | Kohjin Co Ltd | グルタチオン高含有酵母の製造法 |
JP2006042638A (ja) * | 2004-08-02 | 2006-02-16 | Asahi Breweries Ltd | 酵母変異株、グルタチオン高含有酵母の製造方法、その培養物、その分画物、酵母エキスおよびグルタチオン含有飲食品 |
WO2006025295A1 (ja) | 2004-08-30 | 2006-03-09 | Asahi Breweries, Ltd. | マンナンタンパク質を放出する酵母株およびマンナンタンパク質の製造法 |
JP2010073818A (ja) | 2008-09-17 | 2010-04-02 | Canon Inc | 測定装置、露光装置およびデバイス製造方法 |
Non-Patent Citations (4)
Title |
---|
ADAMA, C. ET AL.: "Optimization of biomass and dihydroorotase (DHOase) production by Saccharomyces cerevisiae MNJ3 (pMNJ1)", AFR.J. BIOTECHNOL., vol. 8, no. 1, 2009, pages 37 - 41, XP008161440 * |
LIANG, G. ET AL.: "Elevated glutathione production by adding precursor amino acids coupled with ATP in high cell density cultivation of Candida utilis", J.APPL. MICROBIOL., vol. 105, no. 5, 2008, pages 1432 - 1440, XP008161439 * |
NIELSEN, M.K. ET AL.: "The effect of citric acid and pH on growth and metabolism of anaerobic Saccharomyces cerevisiae and Zygosaccharomyces bailii cultures", FOOD MICROBIOL., vol. 24, no. 1, 2007, pages 101 - 105, XP005593084 * |
TITZE, ANALYTICAL BIOCHEMISTRY, vol. 27, 1969, pages 502 |
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Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2013146465A1 (ja) * | 2012-03-30 | 2013-10-03 | オリエンタル酵母工業株式会社 | 銅高含有酵母抽出物及びその製造方法、並びに、食品、及び野菜の緑色保持復元剤 |
CN104219967A (zh) * | 2012-03-30 | 2014-12-17 | 东方酵母工业株式会社 | 含有高量铜的酵母萃取物及其制造方法以及食品及蔬菜的绿色保持复元剂 |
JPWO2013146465A1 (ja) * | 2012-03-30 | 2015-12-10 | オリエンタル酵母工業株式会社 | 銅高含有酵母抽出物及びその製造方法、並びに、食品、及び野菜の緑色保持復元剤 |
CN102776258A (zh) * | 2012-08-13 | 2012-11-14 | 苏州大学 | 一种s-腺苷-l-蛋氨酸和谷胱甘肽联产发酵方法 |
WO2015151867A1 (ja) * | 2014-03-31 | 2015-10-08 | 興人ライフサイエンス株式会社 | グルタチオンを含む酵母抽出物のメラニン産生抑制剤としての利用 |
JPWO2015151867A1 (ja) * | 2014-03-31 | 2017-04-13 | 興人ライフサイエンス株式会社 | グルタチオンを含む酵母抽出物のメラニン産生抑制剤としての利用 |
JP2019137655A (ja) * | 2018-02-14 | 2019-08-22 | 共栄化学工業株式会社 | 皮膚外用剤 |
WO2020050273A1 (ja) * | 2018-09-07 | 2020-03-12 | 株式会社カネカ | グルタチオンの製造方法 |
CN112639117A (zh) * | 2018-09-07 | 2021-04-09 | 株式会社钟化 | 谷胱甘肽的制造方法 |
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JP5883780B2 (ja) | 2016-03-15 |
JPWO2011118807A1 (ja) | 2013-07-04 |
EP2554656A1 (en) | 2013-02-06 |
EP2554656A4 (en) | 2014-03-26 |
EP2554656B1 (en) | 2018-05-16 |
CN102812119B (zh) | 2015-05-20 |
PL2554656T3 (pl) | 2018-10-31 |
CN102812119A (zh) | 2012-12-05 |
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