WO2006013736A1 - Mutant yeast, method of producing glutathione-rich yeast, culture thereof, fraction thereof, yeast extract and glutathione-containing foods and drinks - Google Patents

Mutant yeast, method of producing glutathione-rich yeast, culture thereof, fraction thereof, yeast extract and glutathione-containing foods and drinks Download PDF

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Publication number
WO2006013736A1
WO2006013736A1 PCT/JP2005/013471 JP2005013471W WO2006013736A1 WO 2006013736 A1 WO2006013736 A1 WO 2006013736A1 JP 2005013471 W JP2005013471 W JP 2005013471W WO 2006013736 A1 WO2006013736 A1 WO 2006013736A1
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Prior art keywords
yeast
dartathione
culture
yeast mutant
mutant
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PCT/JP2005/013471
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French (fr)
Japanese (ja)
Inventor
Kazue Yamamoto
Kaori Yoshimura
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Asahi Breweries, Ltd.
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Priority claimed from JP2004226057A external-priority patent/JP4620404B2/en
Priority claimed from JP2004226059A external-priority patent/JP4620405B2/en
Application filed by Asahi Breweries, Ltd. filed Critical Asahi Breweries, Ltd.
Priority to KR1020077004875A priority Critical patent/KR101167345B1/en
Priority to BRPI0513617-2A priority patent/BRPI0513617A/en
Publication of WO2006013736A1 publication Critical patent/WO2006013736A1/en

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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
    • 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
    • C12N1/145—Fungal 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
    • C12R2001/645—Fungi ; Processes using fungi

Definitions

  • Yeast mutant method for producing yeast having high glutathione content, culture thereof, fraction thereof, yeast extract and food and drink containing glutathione
  • the present invention relates to a yeast mutant of the genus Candida. More specifically, a mutant strain of Candidutilis that can contain a large amount of reduced or acidic salt type dartathione in the microbial cells, a culture obtained by culturing the mutant strain, a fraction, a yeast extract and the like are used. Related to food and drink. Background art
  • Glutathione is a tripeptide composed of three amino acids: glutamic acid, glycine, and cysteine. It is widely distributed in tissues such as the liver and muscles of yeast and animals, and is detoxified and reduced in acid in vivo. Involved in equilibrium action. Focusing on this physiological activity, daltathion has been widely used as a therapeutic agent for liver diseases such as a therapeutic agent for alcoholic fatty liver.
  • Conventionally known methods for producing yeast having a high content of dartathione include a method of adding an amino acid to a medium (Patent Document 1), a method of restricting zinc ions (Patent Document 2), and a culture temperature lower than usual. There is a method of culturing (Patent Document 3). In addition, there are ethionine 'sulfite resistant strains (Patent Document 4) and polyene antibiotic resistant strains (Patent Document 5) by mutagen treatment.
  • Patent Document 1 Japanese Patent Laid-Open No. 53-94089
  • Patent Document 2 JP-A-1 141591
  • Patent Document 3 JP-A-60-156379
  • Patent Document 4 Japanese Patent Laid-Open No. 59-151894
  • Patent Document 5 Japanese Patent Laid-Open No. 2003-284547
  • an object of the present invention is to provide a fermented mother that can retain a large amount of dartathione in a microbial cell and a dartathione-containing food or drink using the same yeast.
  • yeast mutants having cadmium resistance or macrolide antibiotic resistance are contained in a larger amount of cells than general wild yeast. It was found that dartathione can be contained, and the present invention has been completed.
  • the present invention is as follows.
  • a yeast mutant strain that has resistance to cadmium or macrolide antibiotics and can contain a high concentration of dartathione.
  • yeast mutant according to (1), wherein the yeast mutant having cadmium resistance is Candida utilis ABYC1560 (Accession No. FERM P-20094).
  • the yeast mutant having the resistance to macrolide antibiotics is Candida utilis.
  • the yeast mutant according to (1) which is ABYC1561 (Accession No. FERM P-20095).
  • the yeast mutant strain according to any one of (1) to (5) is aerobically cultured, and dartathione is accumulated at a high concentration in the yeast cell.
  • the yeast mutant according to any one of (1) to (5) is cultured under aerobic conditions. A fraction of the culture containing the resulting dartathione.
  • the novel yeast mutant strain of the present invention can contain daltathione at a high concentration in the microbial cells.
  • the novel yeast mutant strain of the present invention is aerobically cultured and cultivated in the yeast microbial cells. Dalthione can be accumulated at high concentrations.
  • the novel yeast mutant of the present invention can be cultured under aerobic conditions to obtain a culture or a fraction containing a high concentration of dartathione.
  • a novel yeast mutant strain of the present invention can be cultured under aerobic conditions to obtain a yeast extract containing daltathione at a high concentration.
  • a culture obtained by culturing the novel yeast mutant of the present invention under aerobic conditions, a fraction of the culture containing dartathione, or the culture or fraction obtained by heat treatment It can be set as a dartathione containing food / beverage product by containing.
  • the novel yeast mutant of the present invention is a yeast mutant having cadmium resistance or macrolide antibiotic resistance and capable of containing dartathione at a high concentration.
  • the cadmium-resistant yeast mutant of the present invention is obtained by selecting Candida utilis 15-10 or the like as a parent strain and selecting a mutant strain having enhanced heavy metal resistance than the parent strain by mutation treatment. It is done.
  • mutagenesis is performed using mutagens such as ultraviolet rays, ionizing radiation, nitrous acid, nitrosoguanidine, and ethenyl methanesulfonate (hereinafter abbreviated as "EMS").
  • mutagens such as ultraviolet rays, ionizing radiation, nitrous acid, nitrosoguanidine, and ethenyl methanesulfonate (hereinafter abbreviated as "EMS").
  • EMS ethenyl methanesulfonate
  • the amount of dartathione contained in the cells of the selected mutant strain is measured, a mutant strain having an increased amount of dartathione is further selected, and a mutant strain that can contain daltathione at a high concentration is searched.
  • a heavy metal used at this time cadmium, mercury, or the like is used.
  • a novel mutant, Candidutilis ABYC1560 was obtained.
  • the mutant Candida utilis ABYC1560 of the present invention contains 0.
  • dartathione in the present invention means the total amount of dartathione of reduced type and acid type dartathione.
  • the general bacteriological properties of the novel mutant Candida utilis ABYC1560 strain of the present invention are exactly the same as the general bacteriological properties of Candida utilis except for cadmium resistance.
  • the novel mutant Candida utilis ABYC1560 strain of the present invention is designated as “Accession number FERM”.
  • the yeast mutant of the present invention may be any one as long as it has resistance to macrolide antibiotics and can contain a large amount of oxidized and reduced darthathione in the microbial cells.
  • Candida utilis ABYC 1561 obtained by mutation treatment using Candida utilis ABYC1560 as a parent strain can be mentioned.
  • the macrolide antibiotic is an antibiotic having macrocyclic ratatones, and examples of the macrolide antibiotic of the present invention include rapamycin, reinamycin, and lankacidin C.
  • yeast mutant strain having resistance to the macrolide antibiotic of the present invention specifically, mutagen such as ultraviolet ray, ionizing radiation, nitrous acid, nitrosoguanidine, EMS, etc. is used.
  • a strain that has grown on an agar medium containing an antibiotic is selected.
  • the amount of dartathione contained in the cells of the selected mutant strain is measured, a mutant strain with an enhanced amount of dartathione is further selected, and a mutant strain that can contain daltathione at a high concentration is searched.
  • a novel mutant Candida utilis ABYC1561 was obtained.
  • Mutant Candida utilis ABY having resistance to macrolide constituents of the present invention C1561 accumulates 0.5 wt% or more of dartathione per dry weight in the microbial cells.
  • dartathione in the present invention means the total amount of dartathione of reduced type and acid type dartathione.
  • novel mutant Candida utilis ABYC1561 strain of the present invention are exactly the same as the general bacteriological properties of Candida utilis except for resistance to macrolide antibiotics. It is.
  • the novel mutant Candida utilis ABYC1561 strain of the present invention is designated as ⁇ Accession No.
  • the present invention further provides a method for producing yeast having a high content of dartathione, wherein the mutant strain is aerobically cultured.
  • the present invention further provides a method for producing a yeast having a high content of dartathione, wherein the yeast mutant strain is aerobically cultured.
  • the cadmium-resistant mutant strain or macrolide antibiotic-resistant mutant strain obtained by the above-described method may be aerobically cultured in a medium containing a carbon source, a nitrogen source, an inorganic salt, and the like. .
  • the medium composition of these strains is one or more selected from the group consisting of glucose, sucrose, acetic acid, ethanol, molasses, and sulfite pulp waste liquor, which are used as a carbon source for the cultivation of ordinary microorganisms.
  • nitrogen source inorganic salts such as urea, ammonia, ammonium sulfate, ammonium chloride or ammonium phosphate, and corn staple (CSL), casein, yeast extract or One or more selected from the group consisting of nitrogen-containing organic substances such as peptone are used.
  • phosphoric acid component, potassium component, magnesium component may be added to the medium, such as normal calcium phosphate, phosphate, potassium chloride, potassium hydroxide, magnesium sulfate, magnesium hydrochloride, etc.
  • Industrial raw material of inorganic salts such as zinc, copper, manganese, and iron ions may be used.
  • vitamins and nucleic acid-related substances may be added.
  • the culture format may be any of batch culture, fed-batch culture or continuous culture, but industrially fed-batch culture or continuous culture is employed.
  • the culture temperature may be in accordance with general yeast culture conditions. For example, 20 to 40 ° C, preferably 25 to 35 ° C is preferable.
  • the pH is 3.5 to 8.0, particularly 4.0 to 6.0.
  • a fraction containing glutathione may be obtained from the strength of the culture that can produce a culture containing a high concentration of dalutathione in yeast.
  • any method can be used as long as it is usually performed! Can be mentioned.
  • by loading the obtained extract on a carrier it can be concentrated to a fraction containing daltathione at a high concentration.
  • a yeast extract may be prepared from the culture cultured by the above method.
  • any method may be used as long as it is a conventional method, but an autolysis method, an enzymatic decomposition method, an alkali extraction method, or the like is industrially employed.
  • dry yeast cells may be prepared from the culture cultured by the above method!
  • any method may be used as long as it is a conventional method, but industrially, freeze drying method, spray drying method, drum drying method, etc. Is adopted.
  • the present invention relates to a culture obtained by culturing the yeast mutant under aerobic conditions, and a food or drink containing a fraction of the culture containing glutathione.
  • These foods and drinks may be any foods and drinks to which dry yeast or yeast extract can be added.
  • a culture obtained by culturing the yeast mutant strain under aerobic conditions, or a fraction of the culture containing daltathione. May be added.
  • the cell weight was measured from the weight after the culture solution was washed twice with a centrifuge and then dried at 105 ° C for 5 hours.
  • Total dartathio combined with reduced and oxidized dartathione Quantification was measured according to the method of Titze et al. (“Analytical Biochemistry”, Vol. 27, page 502, 1969).
  • the total amount of dartathione (% / ⁇ ) in the dry cells was calculated by dividing the total amount of dartathione obtained by the dry cell weight, and was used as a percentage.
  • Candidutilis was cultured until the logarithmic growth phase in a test tube containing YPD medium (glucose 2%, polypeptone 2%, yeastex 1%). Thereafter, the cells were collected and subjected to mutation treatment using EMS according to a conventional method. Mutation treatment was performed under conditions that resulted in a death rate of approximately 70%.
  • the 15-10 strain subjected to the mutation treatment as described above was inoculated into YP D medium containing 0.2 mg / ml of cadmium and statically cultured at 30 ° C for 48 hours. As a result, 200 cadmium-resistant mutant strains were isolated. With respect to these cadmium-resistant mutant strains, the total amount of dartathione was measured by the above-mentioned method, and ABYC1560 strain (accession number FERM P-20094) in which the amount of dartathione in the cells was increased was obtained.
  • P-20094 and its parent strain 15-10 were precultured in YPD medium for 18 hours, then inoculated into YPD medium, and main culture was performed at 30 ° C for 18 hours.
  • the cultured cells were collected by centrifugation, washed twice with water, boiled in a boiling water bath for 5 minutes, extracted from the cells, and the total amount of dartathione was measured according to the method described above.
  • the total amount of dartathione in dry cells is 0.52 wt% in the parent strain (15-10), whereas the cadmium resistant strain (ABYC1560 strain (accession number FE RM
  • Candidutilis was cultured until the logarithmic growth phase in a test tube containing YPD medium (glucose 2%, polypeptone 2%, yeastex 1%). The cells were collected and subjected to mutation treatment using EMS according to a conventional method. Mutation treatment was performed under the condition that the death rate was about 70%.
  • the ABYC1560 strain which was mutated as described above, contained 1 ⁇ g / ml rapamycin.
  • the inoculated YPD agar medium was incubated at 30 ° C for 48 hours. As a result, 745 rapamycin resistant mutants were isolated. With respect to these resistant strains, the total amount of dalutathione was measured by the method described above, and ABYC1561 strain (Accession No. FERM P-20095) with an increased amount of intracellular dalutathione was obtained.
  • the ABYC 1561 strain that acquired rapamycin resistance and its parent strain ABYC 1560 strain were pre-cultured in YP D medium for 18 hours, then inoculated into YPD medium, and main culture was performed at 30 ° C for 18 hours.
  • the cultured cells were collected by centrifugation, washed twice with water, boiled in a boiling water bath for 5 minutes, extracted from the cells, and the total amount of dartathione was measured according to the method described above.
  • the total amount of dartathione in the dried cells is 1.00% in the parent strain (ABYC1560), while the rapamycin metabolite (ABYC1561 (accession number FERM
  • the present invention has industrial applicability in the following points.
  • the novel yeast mutant of the present invention can contain daltathione at a high concentration in the microbial cells, and the novel yeast mutant of the present invention is aerobically cultured in the yeast microbial cells. Dalthione can be accumulated at high concentrations.
  • the novel yeast mutant of the present invention can be cultured under aerobic conditions to obtain a culture or fraction containing a high concentration of dartathione.
  • a novel yeast mutant of the present invention can be cultured under aerobic conditions to obtain a yeast extract containing daltathione at a high concentration.
  • a culture obtained by culturing the novel yeast mutant of the present invention under aerobic conditions, a fraction of the culture containing dartathione, or the culture or fraction obtained by heat treatment It can be set as a dartathione containing food / beverage product by containing.

Abstract

It is intended to provide a yeast capable of holding a large amount of glutathione in its cell. A mutant yeast which is tolerant to cadmium or macrolide antibiotics and can contain a large amount of glutathione.

Description

明 細 書  Specification
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  Yeast mutant, method for producing yeast having high glutathione content, culture thereof, fraction thereof, yeast extract and food and drink containing glutathione
技術分野  Technical field
[0001] 本発明はキャンディダ属の酵母変異株に関する。更に詳しくは、還元型又は酸ィ匕 型ダルタチオンを菌体内に多量に含有できるキャンディダ ュティリスの変異株、当 該変異株を培養して得られる培養物、分画物、酵母エキスおよびこれらを利用した飲 食品に関する。 背景技術  [0001] The present invention relates to a yeast mutant of the genus Candida. More specifically, a mutant strain of Candidutilis that can contain a large amount of reduced or acidic salt type dartathione in the microbial cells, a culture obtained by culturing the mutant strain, a fraction, a yeast extract and the like are used. Related to food and drink. Background art
[0002] グルタチオンはグルタミン酸、グリシン、システィンの 3つのアミノ酸から構成されるト リペプチドであって、酵母や動物の肝臓、筋肉などの組織中に広く分布し、生体内で 解毒作用および酸ィ匕還元平衡作用に関与している。この生理活性に着目して、ダル タチオンはアルコール性脂肪肝の治療薬など肝臓疾患治療薬として広く用いられて きた。  [0002] Glutathione is a tripeptide composed of three amino acids: glutamic acid, glycine, and cysteine. It is widely distributed in tissues such as the liver and muscles of yeast and animals, and is detoxified and reduced in acid in vivo. Involved in equilibrium action. Focusing on this physiological activity, daltathion has been widely used as a therapeutic agent for liver diseases such as a therapeutic agent for alcoholic fatty liver.
[0003] 一方、近年、健康志向の観点より健康機能性食品が脚光を浴びるにつれて、食品 分野でもダルタチオンの生理活性に基づき、同物質含有食品が注目されつつある。  [0003] On the other hand, in recent years, as health functional foods have attracted attention from a health-oriented viewpoint, foods containing the same substance are attracting attention in the food field based on the physiological activity of dartathione.
[0004] ダルタチオンの工業的製造方法としては、酵母力 抽出する方法が一般的である 力 この場合、ダルタチオン含有率の高い酵母の取得が必要条件となる。  [0004] As an industrial production method of dartathione, a method of extracting yeast power is common. In this case, it is necessary to obtain yeast having a high content of dartathione.
[0005] 従来知られているダルタチオン高含有酵母の製造方法としては、培地にアミノ酸添 加する方法 (特許文献 1)、亜鉛イオンを制限する方法 (特許文献 2)、通常よりも低い 培養温度で培養する方法 (特許文献 3)などがある。また、また、突然変異剤処理によ り、ェチォニン'亜硫酸塩耐性株 (特許文献 4)、ポリェン系抗生物質耐性株 (特許文 献 5)などがある。  [0005] Conventionally known methods for producing yeast having a high content of dartathione include a method of adding an amino acid to a medium (Patent Document 1), a method of restricting zinc ions (Patent Document 2), and a culture temperature lower than usual. There is a method of culturing (Patent Document 3). In addition, there are ethionine 'sulfite resistant strains (Patent Document 4) and polyene antibiotic resistant strains (Patent Document 5) by mutagen treatment.
[0006] し力しながら、工業的に安価にダルタチオンを取得するためには、更なるダルタチ オン高含有酵母が望まれる。  [0006] However, in order to obtain dartathione at an industrially low cost, a further yeast with a high daltathione content is desired.
特許文献 1：特開昭 53— 94089号公報  Patent Document 1: Japanese Patent Laid-Open No. 53-94089
特許文献 2 :特開平 1 141591号公報 特許文献 3 :特開昭 60— 156379号公報 Patent Document 2: JP-A-1 141591 Patent Document 3: JP-A-60-156379
特許文献 4:特開昭 59— 151894号公報  Patent Document 4: Japanese Patent Laid-Open No. 59-151894
特許文献 5：特開 2003 - 284547公報  Patent Document 5: Japanese Patent Laid-Open No. 2003-284547
発明の開示  Disclosure of the invention
発明が解決しょうとする課題  Problems to be solved by the invention
[0007] 本発明は、上記の技術背景の下に、菌体内に多量のダルタチオンを保持しうる酵 母および同酵母を利用したダルタチオン含有飲食品を提供することを課題とする。 課題を解決するための手段 [0007] Based on the above technical background, an object of the present invention is to provide a fermented mother that can retain a large amount of dartathione in a microbial cell and a dartathione-containing food or drink using the same yeast. Means for solving the problem
[0008] 本発明者らは上記課題を解決するために鋭意検討を行った結果、カドミウム耐性 又はマクロライド系抗生物質耐性を有する酵母変異株が、一般的な野生酵母よりも 多量の菌体内にダルタチオンを含有しうることを見出し、本発明を完成するに至った [0008] As a result of intensive studies to solve the above problems, the present inventors have found that yeast mutants having cadmium resistance or macrolide antibiotic resistance are contained in a larger amount of cells than general wild yeast. It was found that dartathione can be contained, and the present invention has been completed.
[0009] すなわち、本発明は、以下のとおりである。 That is, the present invention is as follows.
(1) カドミウム耐性又はマクロライド系抗生物質耐性を有し、ダルタチオンを高濃度 で含有することが可能な酵母変異株。  (1) A yeast mutant strain that has resistance to cadmium or macrolide antibiotics and can contain a high concentration of dartathione.
(2) 前記酵母変異株のダルタチオン菌体内含有量が 0.3wt%以上である（1)記載 の酵母変異株。  (2) The yeast mutant according to (1), wherein the content of the yeast mutant in dartathione is 0.3 wt% or more.
(3) 前記カドミウム耐性を有する酵母変異株がキャンディダ'ュティリス ABYC1560 (受託番号 FERM P-20094)である（1)に記載の酵母変異株。  (3) The yeast mutant according to (1), wherein the yeast mutant having cadmium resistance is Candida utilis ABYC1560 (Accession No. FERM P-20094).
(4) 前記マクロライド系抗生物質がラバマイシンである（1)に記載の酵母変異株。（ 5) 前記マクロライド系抗生物質耐性を有する酵母変異株がキャンディダ ·ュティリス (4) The yeast mutant according to (1), wherein the macrolide antibiotic is ravamycin. (5) The yeast mutant having the resistance to macrolide antibiotics is Candida utilis.
ABYC1561 (受託番号 FERM P-20095)である（1)に記載の酵母変異株。The yeast mutant according to (1), which is ABYC1561 (Accession No. FERM P-20095).
(6) (1)ないし (5)のいずれか 1項に記載の酵母変異株を好気的に培養して、当該 酵母菌体内にダルタチオンを高濃度に蓄積させることを特徴とする、ダルタチオン高 含有酵母の製造方法。 (6) The yeast mutant strain according to any one of (1) to (5) is aerobically cultured, and dartathione is accumulated at a high concentration in the yeast cell. A method for producing yeast containing.
(7) (1)ないし (5)のいずれか 1項に記載の酵母変異株を好気的な条件で培養して 得られる培養物。  (7) A culture obtained by culturing the yeast mutant according to any one of (1) to (5) under aerobic conditions.
(8) (1)ないし (5)のいずれか 1項に記載の酵母変異株を好気的な条件で培養して 得られるダルタチオンを含む前記培養物の分画物。 (8) The yeast mutant according to any one of (1) to (5) is cultured under aerobic conditions. A fraction of the culture containing the resulting dartathione.
(9) (1)ないし (5)のいずれか 1項に記載の酵母変異株を好気的な条件で培養して 得られる培養物を用いて製造された酵母エキス。  (9) A yeast extract produced using a culture obtained by culturing the yeast mutant according to any one of (1) to (5) under aerobic conditions.
(10) (1)ないし (5)のいずれか 1項に記載の酵母変異株を好気的な条件で培養し て得られる培養物を用いて製造された乾燥酵母菌体。  (10) A dry yeast cell produced using a culture obtained by culturing the yeast mutant according to any one of (1) to (5) under an aerobic condition.
(11) (1)ないし (5)のいずれか 1項に記載の酵母変異株を好気的な条件で培養し て得られる培養物、ダルタチオンを含む前記培養物の分画物を含有することを特徴 とするダルタチオン含有飲食品。  (11) A culture obtained by culturing the yeast mutant according to any one of (1) to (5) under aerobic conditions, and a fraction of the culture containing daltathione A food and drink containing dartathione, characterized by
発明の効果  The invention's effect
[0010] 本発明の新規な酵母変異株は菌体内にダルタチオンを高濃度で含有することが可 能であり、本発明の新規な酵母変異株を好気的に培養して当該酵母菌体内にダル タチオンを高濃度に蓄積させることができる。  [0010] The novel yeast mutant strain of the present invention can contain daltathione at a high concentration in the microbial cells. The novel yeast mutant strain of the present invention is aerobically cultured and cultivated in the yeast microbial cells. Dalthione can be accumulated at high concentrations.
[0011] 従って、本発明の新規な酵母変異株を好気的な条件で培養して、ダルタチオンを 高濃度に含有する培養物又は分画物を得ることができる。  [0011] Therefore, the novel yeast mutant of the present invention can be cultured under aerobic conditions to obtain a culture or a fraction containing a high concentration of dartathione.
[0012] また、本発明の新規な酵母変異株を好気的な条件で培養して、ダルタチオンを高 濃度に含有する酵母エキスを得ることができる。  [0012] In addition, a novel yeast mutant strain of the present invention can be cultured under aerobic conditions to obtain a yeast extract containing daltathione at a high concentration.
[0013] さらにまた、本発明の新規な酵母変異株を好気的な条件で培養して得られる培養 物、ダルタチオンを含む前記培養物の分画物又は加熱処理した前記培養物もしくは 分画物を含有させることによりダルタチオン含有飲食品とすることができる。  [0013] Furthermore, a culture obtained by culturing the novel yeast mutant of the present invention under aerobic conditions, a fraction of the culture containing dartathione, or the culture or fraction obtained by heat treatment It can be set as a dartathione containing food / beverage product by containing.
発明を実施するための最良の形態  BEST MODE FOR CARRYING OUT THE INVENTION
[0014] 以下、本発明を詳細に説明する。 [0014] Hereinafter, the present invention will be described in detail.
[0015] 本発明の、新規な酵母変異株は、カドミウム耐性又はマクロライド系抗生物質耐性 を有し、ダルタチオンを高濃度で含有することが可能な酵母変異株である。  [0015] The novel yeast mutant of the present invention is a yeast mutant having cadmium resistance or macrolide antibiotic resistance and capable of containing dartathione at a high concentration.
[0016] また、本発明のカドミウム耐性を有する酵母変異株はキャンディダ'ュティリス 15-10 株 等を親株とし、突然変異処理により親株よりも重金属耐性能の亢進した変異株を 選択することにより得られる。 [0016] Further, the cadmium-resistant yeast mutant of the present invention is obtained by selecting Candida utilis 15-10 or the like as a parent strain and selecting a mutant strain having enhanced heavy metal resistance than the parent strain by mutation treatment. It is done.
[0017] 具体的には、紫外線、電離放射線、亜硝酸、ニトロソグァ-ジン、メタンスルホン酸 ェチル (以下、「EMS」と略称する）などの変異原を用いて突然変異処理を行い、カド ミゥム等の重金属を含む培地上で生育が認められた変異株を選択する。 [0017] Specifically, mutagenesis is performed using mutagens such as ultraviolet rays, ionizing radiation, nitrous acid, nitrosoguanidine, and ethenyl methanesulfonate (hereinafter abbreviated as "EMS"). Select a mutant strain that has grown on a medium containing heavy metals such as Myum.
[0018] 選択した変異株の菌体中に含まれるダルタチオン量を測定し、ダルタチオン量が増 強した変異株をさらに選択し、ダルタチオンを高濃度で含有しうる変異株を探索する 。この際使用する重金属としては、カドミウム、水銀などが用いられる。このようにして 本発明では、新規な変異株であるキャンディダ ·ュティリス ABYC1560が得られた。  [0018] The amount of dartathione contained in the cells of the selected mutant strain is measured, a mutant strain having an increased amount of dartathione is further selected, and a mutant strain that can contain daltathione at a high concentration is searched. As the heavy metal used at this time, cadmium, mercury, or the like is used. Thus, in the present invention, a novel mutant, Candidutilis ABYC1560, was obtained.
[0019] 本発明の変異株キャンディダ'ュティリス ABYC1560は菌体中に、乾燥重量当たり 0.  [0019] The mutant Candida utilis ABYC1560 of the present invention contains 0.
3wt%以上のダルタチオンを蓄積する。本発明におけるダルタチオンとは、還元型お よび酸ィ匕型のダルタチオンの総ダルタチオン量を意味するものである。  Accumulate 3 wt% or more of dartathione. The term dartathione in the present invention means the total amount of dartathione of reduced type and acid type dartathione.
[0020] また、本発明の新規変異キャンディダ*ュティリス ABYC1560株の一般的菌学的性 質は、カドミウム耐性を除いてはキャンディダ ·ュティリスの一般的菌学的性質と全く 同一である。本発明の新規変異キャンディダ*ュティリス ABYC1560株は、「受託番号 FERM  [0020] In addition, the general bacteriological properties of the novel mutant Candida utilis ABYC1560 strain of the present invention are exactly the same as the general bacteriological properties of Candida utilis except for cadmium resistance. The novel mutant Candida utilis ABYC1560 strain of the present invention is designated as “Accession number FERM”.
P-20094Jとして受託された。  Commissioned as P-20094J.
[0021] また、本発明の酵母変異株はマクロライド系抗生物質耐性を有し、更には酸化型お よび還元型ダルタチオンを多量に菌体内に含有できるものであれば、いずれであつ てもよい。具体的には、キャンディダ 'ュティリス ABYC1560を親株とし、変異処理によ つて得られたキャンディダ ·ュティリス ABYC 1561などが挙げられる。  [0021] Furthermore, the yeast mutant of the present invention may be any one as long as it has resistance to macrolide antibiotics and can contain a large amount of oxidized and reduced darthathione in the microbial cells. . Specifically, Candida utilis ABYC 1561 obtained by mutation treatment using Candida utilis ABYC1560 as a parent strain can be mentioned.
[0022] マクロライド系抗生物質とは、大環状ラタトンを有する抗生物質であり、本発明のマ クロライド系抗生物質として、例えばラパマイシン、レイナマイシン、ランカシディン C等 を挙げることができる。  [0022] The macrolide antibiotic is an antibiotic having macrocyclic ratatones, and examples of the macrolide antibiotic of the present invention include rapamycin, reinamycin, and lankacidin C.
[0023] 本発明のマクロライド系抗生物質耐性を有する酵母変異株を得るには、具体的に は、紫外線、電離放射線、亜硝酸、ニトロソグァ-ジン、 EMSなどの変異原を用い、 マクロライド系抗生物質を含む寒天培地上で生育が認められた株を選択する。選択 した変異株の菌体中に含まれるダルタチオン量を測定し、ダルタチオン量が増強した 変異株をさらに選択し、ダルタチオンを高濃度で含有しうる変異株を探索する。このよ うにして本発明では、新規な変異株であるキャンディダ'ュティリス ABYC1561が得ら れた。  [0023] In order to obtain the yeast mutant strain having resistance to the macrolide antibiotic of the present invention, specifically, mutagen such as ultraviolet ray, ionizing radiation, nitrous acid, nitrosoguanidine, EMS, etc. is used. A strain that has grown on an agar medium containing an antibiotic is selected. The amount of dartathione contained in the cells of the selected mutant strain is measured, a mutant strain with an enhanced amount of dartathione is further selected, and a mutant strain that can contain daltathione at a high concentration is searched. Thus, in the present invention, a novel mutant Candida utilis ABYC1561 was obtained.
[0024] 本発明のマクロライド系構成物質耐性を有する変異株キャンディダ ·ュティリス ABY C1561は菌体中に、乾燥重量当たり 0.5wt%以上のダルタチオンを蓄積する。本発明 におけるダルタチオンとは、還元型および酸ィヒ型のダルタチオンの総ダルタチオン量 を意味するものである。 [0024] Mutant Candida utilis ABY having resistance to macrolide constituents of the present invention C1561 accumulates 0.5 wt% or more of dartathione per dry weight in the microbial cells. The term dartathione in the present invention means the total amount of dartathione of reduced type and acid type dartathione.
[0025] さらにまた、本発明の新規変異キャンディダ'ュティリス ABYC1561株の一般的菌学 的性質は、マクロライド系抗生物質耐性を除いてはキャンディダ'ュティリスの一般的 菌学的性質と全く同一である。本発明の新規変異キャンディダ 'ュティリス ABYC1561 株は、「受託番号 FERM  [0025] Furthermore, the general bacteriological properties of the novel mutant Candida utilis ABYC1561 strain of the present invention are exactly the same as the general bacteriological properties of Candida utilis except for resistance to macrolide antibiotics. It is. The novel mutant Candida utilis ABYC1561 strain of the present invention is designated as `` Accession No.
P-20095Jとして受託された。  Commissioned as P-20095J.
[0026] 本発明では、更に、変異株を好気的に培養するダルタチオン高含有酵母の製造方 法が提供される。  [0026] The present invention further provides a method for producing yeast having a high content of dartathione, wherein the mutant strain is aerobically cultured.
[0027] 本発明では、更に、上記酵母変異株を好気的に培養するダルタチオン高含有酵母 の製造方法が提供される。本発明を実施するには、前述の方法で得られたカドミウム 耐性変異株又はマクロライド系抗生物質耐性変異株を炭素源、窒素源及び無機塩 等を含む培地で好気的に培養すればよい。  [0027] The present invention further provides a method for producing a yeast having a high content of dartathione, wherein the yeast mutant strain is aerobically cultured. In order to carry out the present invention, the cadmium-resistant mutant strain or macrolide antibiotic-resistant mutant strain obtained by the above-described method may be aerobically cultured in a medium containing a carbon source, a nitrogen source, an inorganic salt, and the like. .
[0028] これら菌株の培地組成としては、炭素源として通常の微生物の培養に利用されるグ ルコース、蔗糖、酢酸、エタノール、糖蜜および亜硫酸パルプ廃液等力 なる群より 選ばれる 1種又は 2種以上が用いられ、窒素源としては、尿素、アンモニア、硫酸アン モ-ゥム、塩化アンモ-ゥムもしくはリン酸アンモ-ゥム等の無機塩、およびコーンス ティプリカ一（CSL)、カゼイン、酵母エキスもしくはペプトン等の含窒素有機物等から なる群より選ばれる 1種又は 2種以上が使用される。更に、リン酸成分、カリウム成分、 マグネシウム成分を培地に添加してもよぐこれらとしては、過リン酸石灰、リン安、塩 化カリウム、水酸ィ匕カリウム、硫酸マグネシウム、塩酸マグネシウム等の通常の工業用 原料でよい。その他、亜鉛、銅、マンガン、鉄イオン等の無機塩を使用してもよい。そ の他、ビタミン、核酸関連物質等を添加しても良い。  [0028] The medium composition of these strains is one or more selected from the group consisting of glucose, sucrose, acetic acid, ethanol, molasses, and sulfite pulp waste liquor, which are used as a carbon source for the cultivation of ordinary microorganisms. As the nitrogen source, inorganic salts such as urea, ammonia, ammonium sulfate, ammonium chloride or ammonium phosphate, and corn staple (CSL), casein, yeast extract or One or more selected from the group consisting of nitrogen-containing organic substances such as peptone are used. In addition, phosphoric acid component, potassium component, magnesium component may be added to the medium, such as normal calcium phosphate, phosphate, potassium chloride, potassium hydroxide, magnesium sulfate, magnesium hydrochloride, etc. Industrial raw material of In addition, inorganic salts such as zinc, copper, manganese, and iron ions may be used. In addition, vitamins and nucleic acid-related substances may be added.
[0029] 培養形式としては、回分培養、流加培養又は連続培養の 、ずれでもよ ヽが、工業 的には流加培養又は連続培養が採用される。  [0029] The culture format may be any of batch culture, fed-batch culture or continuous culture, but industrially fed-batch culture or continuous culture is employed.
[0030] 培養温度は、一般的な酵母の培養条件に従えばよく、例えば 20〜40°C、好ましくは 25〜35°Cがよぐ pHは 3.5〜8.0、特に 4.0〜6.0が望ましい。 [0031] 本発明の方法により高濃度ダルタチオンを酵母菌体内に含有する培養物が得られ る力 培養物力ゝらグルタチオンを含有する分画物を得てもよい。培養物からダルタチ オンを含有する分画物を分画する方法としては、通常行われて 、る方法であれば!/、 ずれの方法でもよいが、熱水抽出、菌体破砕による抽出等を挙げることができる。ま た、得られた抽出物を担体に担持させることにより、ダルタチオンを高濃度に含む画 分に濃縮等することが可能となる。 [0030] The culture temperature may be in accordance with general yeast culture conditions. For example, 20 to 40 ° C, preferably 25 to 35 ° C is preferable. The pH is 3.5 to 8.0, particularly 4.0 to 6.0. [0031] According to the method of the present invention, a fraction containing glutathione may be obtained from the strength of the culture that can produce a culture containing a high concentration of dalutathione in yeast. As a method for fractionating a fraction containing dartathione from a culture, any method can be used as long as it is usually performed! Can be mentioned. In addition, by loading the obtained extract on a carrier, it can be concentrated to a fraction containing daltathione at a high concentration.
[0032] また、上記方法により培養した培養物から酵母エキスを調製してもよい。酵母エキス を調製する方法としては、通常行われて 、る方法であれば 、ずれの方法であっても よいが、自己消化法、酵素分解法あるいはアルカリ抽出法などが工業的に採用され る。  [0032] In addition, a yeast extract may be prepared from the culture cultured by the above method. As a method for preparing a yeast extract, any method may be used as long as it is a conventional method, but an autolysis method, an enzymatic decomposition method, an alkali extraction method, or the like is industrially employed.
[0033] また、上記方法により培養した培養物から乾燥酵母菌体を調製してもよ!/、。乾燥酵 母菌体を調製する方法としては、通常行われて 、る方法であれば 、ずれの方法であ つてもよいが、工業的には、凍結乾燥法、スプレードライ法、ドラムドライ法などが採用 される。  [0033] Alternatively, dry yeast cells may be prepared from the culture cultured by the above method! As a method for preparing dry fermentation mother cells, any method may be used as long as it is a conventional method, but industrially, freeze drying method, spray drying method, drum drying method, etc. Is adopted.
[0034] さらに本発明は、上記酵母変異株を好気的な条件で培養して得られる培養物、グ ルタチオンを含む前記培養物の分画物を含有する飲食品に関するものである。  [0034] Further, the present invention relates to a culture obtained by culturing the yeast mutant under aerobic conditions, and a food or drink containing a fraction of the culture containing glutathione.
[0035] これらの飲食品としては、通常乾燥酵母、酵母エキスを添加しうる飲食品であれば 何れでもよいが、例えばアルコール飲料、清涼飲料、発酵食品調味料、スープ類、 一般食品、菓子類等を挙げることができる。 [0035] These foods and drinks may be any foods and drinks to which dry yeast or yeast extract can be added. For example, alcoholic beverages, soft drinks, fermented food seasonings, soups, general foods, confectionery Etc.
[0036] 本発明の飲食品を製造するには、飲食品の製造工程において、上記酵母変異株 を好気的な条件で培養して得られる培養物、ダルタチオンを含む前記培養物の分画 物を添加すればよい。 [0036] In order to produce the food or drink of the present invention, in the production process of the food or drink, a culture obtained by culturing the yeast mutant strain under aerobic conditions, or a fraction of the culture containing daltathione. May be added.
[0037] 従って、本発明によって、ダルタチオンを高濃度で含む飲食品を効率的に製造す ることがでさる。  [0037] Therefore, according to the present invention, it is possible to efficiently produce a food or drink containing a high concentration of dartathione.
[0038] 以下、本発明を、実施例を挙げて説明する。本発明は、これら実施例に限定される ものではない。  Hereinafter, the present invention will be described with reference to examples. The present invention is not limited to these examples.
[0039] なお、菌体重量の測定は培養液を遠心分離機で 2回水洗後、 105°C、 5時間乾燥さ せた後の重量から求めた。還元型及び酸化型ダルタチオンを合わせた総ダルタチォ ンの定量は Titzeらの方法（「アナリティカル バイオケミストリー」第 27卷、 502ページ、 1969年）に従って測定した。乾燥菌体中の総ダルタチオン量（％ /^)は得られた総 ダルタチオン量を乾燥菌体重量で割ることにより算出して百分率とした。 [0039] The cell weight was measured from the weight after the culture solution was washed twice with a centrifuge and then dried at 105 ° C for 5 hours. Total dartathio combined with reduced and oxidized dartathione Quantification was measured according to the method of Titze et al. (“Analytical Biochemistry”, Vol. 27, page 502, 1969). The total amount of dartathione (% / ^) in the dry cells was calculated by dividing the total amount of dartathione obtained by the dry cell weight, and was used as a percentage.
実施例 1  Example 1
[0040] キャンディダ ュティリスを YPD培地（グルコース 2%、ポリペプトン 2%、イーストェキ ス 1%)を含む試験管で対数増殖期まで培養した。その後、この菌体を回収し、常法 に従い EMSを用いて変異処理を行った。変異処理は死滅率約 70%になるような条 件で行った。  [0040] Candidutilis was cultured until the logarithmic growth phase in a test tube containing YPD medium (glucose 2%, polypeptone 2%, yeastex 1%). Thereafter, the cells were collected and subjected to mutation treatment using EMS according to a conventional method. Mutation treatment was performed under conditions that resulted in a death rate of approximately 70%.
[0041] 上記のようにして変異処理を施した 15-10株を、 0.2mg/mlのカドミウムを含有する YP D培地に植菌し、 30°Cで 48時間静置培養した。その結果、 200株のカドミウム耐性変 異株を単離した。これらカドミウム耐性変異株について、上述の方法で総ダルタチォ ン量の測定を行 、菌体内ダルタチオン量が増強した ABYC1560株（受託番号 FERM P-20094)を取得した。  [0041] The 15-10 strain subjected to the mutation treatment as described above was inoculated into YP D medium containing 0.2 mg / ml of cadmium and statically cultured at 30 ° C for 48 hours. As a result, 200 cadmium-resistant mutant strains were isolated. With respect to these cadmium-resistant mutant strains, the total amount of dartathione was measured by the above-mentioned method, and ABYC1560 strain (accession number FERM P-20094) in which the amount of dartathione in the cells was increased was obtained.
実施例 2  Example 2
[0042] カドミウム耐性を獲得した ABYC 1560株（受託番号 FERM  [0042] ABYC 1560 strain that has acquired cadmium resistance (Accession No. FERM)
P-20094)及びその親株である 15-10株を YPD培地で 18時間前培養した後、 YPD培地 に植菌し、 18時間 30°Cで本培養を行った。培養後の菌体を遠心分離で回収した後 、 2回水洗後、沸騰水浴中で 5分煮沸し、菌体内ダルタチオンを抽出し、上述の方法 に従って総ダルタチオン量の測定を実施した。乾燥菌体中の総ダルタチオン量は親 株（15-10)が 0.52wt%であるのに対し、カドミウム耐性株 (ABYC1560株（受託番号 FE RM  P-20094) and its parent strain 15-10 were precultured in YPD medium for 18 hours, then inoculated into YPD medium, and main culture was performed at 30 ° C for 18 hours. The cultured cells were collected by centrifugation, washed twice with water, boiled in a boiling water bath for 5 minutes, extracted from the cells, and the total amount of dartathione was measured according to the method described above. The total amount of dartathione in dry cells is 0.52 wt% in the parent strain (15-10), whereas the cadmium resistant strain (ABYC1560 strain (accession number FE RM
P- 20094) )では 1.00wt%であった。  In P-20094)), it was 1.00wt%.
実施例 3  Example 3
[0043] キャンディダ ュティリスを YPD培地（グルコース 2%、ポリペプトン 2%、イーストェキ ス 1%)を含む試験管で対数増殖期まで培養した。この菌体を回収し、常法に従い E MSを用いて変異処理を行った。変異処理は死滅率約 70%になるような条件で行つ た。  [0043] Candidutilis was cultured until the logarithmic growth phase in a test tube containing YPD medium (glucose 2%, polypeptone 2%, yeastex 1%). The cells were collected and subjected to mutation treatment using EMS according to a conventional method. Mutation treatment was performed under the condition that the death rate was about 70%.
[0044] 上記のようにして変異処理を実施した ABYC1560株を 1 μ g/mlのラパマイシンを含 有する YPD寒天培地に植菌し、 30°Cで 48時間静置培養した。結果、 745株のラパマイ シン耐性変異株を単離した。これら耐性株について、上述の方法で総ダルタチオン 量の測定を行 、菌体内ダルタチオン量が増強した ABYC1561株（受託番号 FERM P-20095)を取得した。 [0044] The ABYC1560 strain, which was mutated as described above, contained 1 μg / ml rapamycin. The inoculated YPD agar medium was incubated at 30 ° C for 48 hours. As a result, 745 rapamycin resistant mutants were isolated. With respect to these resistant strains, the total amount of dalutathione was measured by the method described above, and ABYC1561 strain (Accession No. FERM P-20095) with an increased amount of intracellular dalutathione was obtained.
実施例 4  Example 4
[0045] ラパマイシン耐性を獲得した ABYC 1561株 及びその親株である ABYC1560株を YP D培地で 18時間前培養した後、 YPD培地に植菌し、 18時間 30°Cで本培養を行った 。培養後の菌体を遠心分離で回収した後、 2回水洗後、沸騰水浴中で 5分煮沸し、菌 体内ダルタチオンを抽出し、上述の方法に従って総ダルタチオン量の測定を実施し た。乾燥菌体中の総ダルタチオン量は親株 (ABYC1560)が 1.00%であるのに対し、 ラパマイシン而性株（ABYC1561 (受託番号 FERM  [0045] The ABYC 1561 strain that acquired rapamycin resistance and its parent strain ABYC 1560 strain were pre-cultured in YP D medium for 18 hours, then inoculated into YPD medium, and main culture was performed at 30 ° C for 18 hours. The cultured cells were collected by centrifugation, washed twice with water, boiled in a boiling water bath for 5 minutes, extracted from the cells, and the total amount of dartathione was measured according to the method described above. The total amount of dartathione in the dried cells is 1.00% in the parent strain (ABYC1560), while the rapamycin metabolite (ABYC1561 (accession number FERM
P-20095) )では 1.82%であった。  P-20095)) was 1.82%.
産業上の利用可能性  Industrial applicability
[0046] 本発明は、以下の点において、産業上の利用可能性が存在する。 [0046] The present invention has industrial applicability in the following points.
[0047] 本発明の新規な酵母変異株は菌体内にダルタチオンを高濃度で含有することが可 能であり、本発明の新規な酵母変異株を好気的に培養して当該酵母菌体内にダル タチオンを高濃度に蓄積させることができる。 [0047] The novel yeast mutant of the present invention can contain daltathione at a high concentration in the microbial cells, and the novel yeast mutant of the present invention is aerobically cultured in the yeast microbial cells. Dalthione can be accumulated at high concentrations.
[0048] 従って、本発明の新規な酵母変異株を好気的な条件で培養して、ダルタチオンを 高濃度に含有する培養物又は分画物を得ることができる。 [0048] Therefore, the novel yeast mutant of the present invention can be cultured under aerobic conditions to obtain a culture or fraction containing a high concentration of dartathione.
[0049] また、本発明の新規な酵母変異株を好気的な条件で培養して、ダルタチオンを高 濃度に含有する酵母エキスを得ることができる。 [0049] In addition, a novel yeast mutant of the present invention can be cultured under aerobic conditions to obtain a yeast extract containing daltathione at a high concentration.
[0050] さらにまた、本発明の新規な酵母変異株を好気的な条件で培養して得られる培養 物、ダルタチオンを含む前記培養物の分画物又は加熱処理した前記培養物もしくは 分画物を含有させることによりダルタチオン含有飲食品とすることができる。 [0050] Furthermore, a culture obtained by culturing the novel yeast mutant of the present invention under aerobic conditions, a fraction of the culture containing dartathione, or the culture or fraction obtained by heat treatment It can be set as a dartathione containing food / beverage product by containing.

Claims

請求の範囲  The scope of the claims
[I] カドミウム耐性又はマクロライド系抗生物質耐性を有し、ダルタチオンを高濃度で含 有することが可能な酵母変異株。  [I] A yeast mutant strain that is resistant to cadmium or macrolide antibiotics and can contain high concentrations of dartathione.
[2] 前記酵母変異株のダルタチオン菌体内含有量力 S0.3wt%以上である請求項 1記載の 酵母変異株。  [2] The yeast mutant according to claim 1, wherein the content of the yeast mutant in dartathione is S0.3 wt% or more.
[3] 前記カドミウム耐性を有する酵母変異株がキャンディダ'ュティリス ABYC1560 (受託 番号 FERM  [3] The cadmium-resistant yeast mutant is Candida utilis ABYC1560 (accession number FERM
P-20094)である請求項 1に記載の酵母変異株。  The yeast mutant according to claim 1, which is P-20094).
[4] 前記マクロライド系抗生物質カ^パマイシンである請求項 1に記載の酵母変異株。 [4] The yeast mutant according to claim 1, which is the macrolide antibiotic kapamycin.
[5] 前記マクロライド系抗生物質耐性を有する酵母変異株がキャンディダ ·ュティリス AB[5] The yeast mutant having resistance to macrolide antibiotics is Candida utilis AB
YC1561 (受託番号 FERM P-20095)である請求項 1に記載の酵母変異株。 The yeast mutant according to claim 1, which is YC1561 (Accession No. FERM P-20095).
[6] 請求項 1ないし 5のいずれ力 1項に記載の酵母変異株を好気的に培養して、当該酵 母菌体内にダルタチオンを高濃度に蓄積させることを特徴とする、ダルタチオン高含 有酵母の製造方法。 [6] In any one of claims 1 to 5, the yeast mutant according to item 1 is aerobically cultured, and dartathione is accumulated in a high concentration in the yeast. A method for producing yeast.
[7] 請求項 1な!ヽし 5の ヽずれか 1項に記載の酵母変異株を好気的な条件で培養して得 られる培養物。  [7] A culture obtained by culturing the yeast mutant according to claim 1 under aerobic conditions.
[8] 請求項 1な!ヽし 5の ヽずれか 1項に記載の酵母変異株を好気的な条件で培養して得 られるダルタチオンを含む前記培養物の分画物。  [8] A fraction of the culture containing dartathione obtained by culturing the yeast mutant according to claim 1 under aerobic conditions.
[9] 請求項 1な!ヽし 5の ヽずれか 1項に記載の酵母変異株を好気的な条件で培養して得 られる培養物を用いて製造された酵母エキス。 [9] A yeast extract produced using a culture obtained by culturing the yeast mutant according to claim 1 under aerobic conditions.
[10] 請求項 1な!ヽし 5の ヽずれか 1項に記載の酵母変異株を好気的な条件で培養して得 られる培養物を用いて製造された乾燥酵母菌体。 [10] A dry yeast cell produced using a culture obtained by culturing the yeast mutant according to claim 1 under aerobic conditions.
[II] 請求項 1ないし 5のいずれ力 1項に記載の酵母変異株を好気的な条件で培養して得 られる培養物、ダルタチオンを含む前記培養物の分画物を含有することを特徴とする ダルタチオン含有飲食品。  [II] A force obtained by culturing the yeast mutant strain according to any one of claims 1 to 5 under an aerobic condition, comprising a fraction of the culture containing dartathione. Daltathione-containing food and drink.
PCT/JP2005/013471 2004-08-02 2005-07-22 Mutant yeast, method of producing glutathione-rich yeast, culture thereof, fraction thereof, yeast extract and glutathione-containing foods and drinks WO2006013736A1 (en)

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