JPH02295480A - Yeast having high glutathione content and production thereof - Google Patents
Yeast having high glutathione content and production thereofInfo
- Publication number
- JPH02295480A JPH02295480A JP11529589A JP11529589A JPH02295480A JP H02295480 A JPH02295480 A JP H02295480A JP 11529589 A JP11529589 A JP 11529589A JP 11529589 A JP11529589 A JP 11529589A JP H02295480 A JPH02295480 A JP H02295480A
- Authority
- JP
- Japan
- Prior art keywords
- glutathione
- yeast
- producing
- zinc ion
- zinc
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 title claims abstract description 74
- 229960003180 glutathione Drugs 0.000 title claims abstract description 38
- 108010024636 Glutathione Proteins 0.000 title claims abstract description 37
- 240000004808 Saccharomyces cerevisiae Species 0.000 title claims abstract description 25
- 238000004519 manufacturing process Methods 0.000 title claims description 8
- PTFCDOFLOPIGGS-UHFFFAOYSA-N Zinc dication Chemical compound [Zn+2] PTFCDOFLOPIGGS-UHFFFAOYSA-N 0.000 claims abstract description 19
- 238000012258 culturing Methods 0.000 claims abstract description 5
- 241000235070 Saccharomyces Species 0.000 claims abstract description 4
- 238000000034 method Methods 0.000 claims description 3
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 abstract description 21
- 239000002609 medium Substances 0.000 description 14
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 8
- 230000001580 bacterial effect Effects 0.000 description 8
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 6
- 210000004027 cell Anatomy 0.000 description 6
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 5
- 239000008103 glucose Substances 0.000 description 5
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 4
- 150000001413 amino acids Chemical class 0.000 description 4
- 229960002685 biotin Drugs 0.000 description 4
- 235000020958 biotin Nutrition 0.000 description 4
- 239000011616 biotin Substances 0.000 description 4
- 244000005700 microbiome Species 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 235000013343 vitamin Nutrition 0.000 description 4
- 239000011782 vitamin Substances 0.000 description 4
- 229940088594 vitamin Drugs 0.000 description 4
- 229930003231 vitamin Natural products 0.000 description 4
- 150000003722 vitamin derivatives Chemical class 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- 239000004202 carbamide Substances 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 238000000605 extraction Methods 0.000 description 3
- 210000005253 yeast cell Anatomy 0.000 description 3
- KPGXRSRHYNQIFN-UHFFFAOYSA-N 2-oxoglutaric acid Chemical compound OC(=O)CCC(=O)C(O)=O KPGXRSRHYNQIFN-UHFFFAOYSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- JIAARYAFYJHUJI-UHFFFAOYSA-L zinc dichloride Chemical compound [Cl-].[Cl-].[Zn+2] JIAARYAFYJHUJI-UHFFFAOYSA-L 0.000 description 2
- WTLKTXIHIHFSGU-UHFFFAOYSA-N 2-nitrosoguanidine Chemical compound NC(N)=NN=O WTLKTXIHIHFSGU-UHFFFAOYSA-N 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- PLUBXMRUUVWRLT-UHFFFAOYSA-N Ethyl methanesulfonate Chemical compound CCOS(C)(=O)=O PLUBXMRUUVWRLT-UHFFFAOYSA-N 0.000 description 1
- 229930091371 Fructose Natural products 0.000 description 1
- 239000005715 Fructose Substances 0.000 description 1
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 description 1
- CANRESZKMUPMAE-UHFFFAOYSA-L Zinc lactate Chemical compound [Zn+2].CC(O)C([O-])=O.CC(O)C([O-])=O CANRESZKMUPMAE-UHFFFAOYSA-L 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- ZOIORXHNWRGPMV-UHFFFAOYSA-N acetic acid;zinc Chemical compound [Zn].CC(O)=O.CC(O)=O ZOIORXHNWRGPMV-UHFFFAOYSA-N 0.000 description 1
- 238000005273 aeration Methods 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- HWXBTNAVRSUOJR-UHFFFAOYSA-N alpha-hydroxyglutaric acid Natural products OC(=O)C(O)CCC(O)=O HWXBTNAVRSUOJR-UHFFFAOYSA-N 0.000 description 1
- 229940009533 alpha-ketoglutaric acid Drugs 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000001784 detoxification Methods 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 235000019264 food flavour enhancer Nutrition 0.000 description 1
- 235000013376 functional food Nutrition 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 229930195733 hydrocarbon Natural products 0.000 description 1
- 150000002430 hydrocarbons Chemical class 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000004811 liquid chromatography Methods 0.000 description 1
- 230000003908 liver function Effects 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 239000003471 mutagenic agent Substances 0.000 description 1
- 231100000707 mutagenic chemical Toxicity 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 235000005985 organic acids Nutrition 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 238000001308 synthesis method Methods 0.000 description 1
- 238000010189 synthetic method Methods 0.000 description 1
- 238000003809 water extraction Methods 0.000 description 1
- 239000011701 zinc Substances 0.000 description 1
- 229910052725 zinc Inorganic materials 0.000 description 1
- 239000004246 zinc acetate Substances 0.000 description 1
- 235000013904 zinc acetate Nutrition 0.000 description 1
- 239000011592 zinc chloride Substances 0.000 description 1
- 235000005074 zinc chloride Nutrition 0.000 description 1
- 150000003752 zinc compounds Chemical class 0.000 description 1
- 239000011576 zinc lactate Substances 0.000 description 1
- 229940050168 zinc lactate Drugs 0.000 description 1
- 235000000193 zinc lactate Nutrition 0.000 description 1
- LRXTYHSAJDENHV-UHFFFAOYSA-H zinc phosphate Chemical compound [Zn+2].[Zn+2].[Zn+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O LRXTYHSAJDENHV-UHFFFAOYSA-H 0.000 description 1
- 229910000165 zinc phosphate Inorganic materials 0.000 description 1
- XOOUIPVCVHRTMJ-UHFFFAOYSA-L zinc stearate Chemical compound [Zn+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O XOOUIPVCVHRTMJ-UHFFFAOYSA-L 0.000 description 1
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 description 1
- 229960001763 zinc sulfate Drugs 0.000 description 1
- 229910000368 zinc sulfate Inorganic materials 0.000 description 1
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
Description
【発明の詳細な説明】
〔産業上の利用分野〕
本発明はサツ力ロミセス属に属する酵母より亜鉛イオン
耐性を獲得した変異株を好気的に培養し、該菌体中に蓄
量のグルタチオンを生成蓄積する酵母の製造方法及びグ
ルタチオン高含有酵母に関するものである。[Detailed Description of the Invention] [Industrial Application Field] The present invention involves aerobically cultivating a mutant strain of yeast belonging to the genus Satsuromyces that has acquired resistance to zinc ions, and culturing a large amount of glutathione in the bacterial cells. The present invention relates to a method for producing yeast that produces and accumulates glutathione, and to a yeast with a high glutathione content.
グルタチオンは、肝臓機能回復作用や解毒作用などの生
理作用を有する。また、ごく味増強物質等として食品に
も利用される．
〔従来技術〕
グルタチオンの工業的製造法としては、合成法、及びバ
ン酵母・ビール酵母菌体からの抽出法が採用されている
。しかし、合成法によるときはＤＬ−グルタチオンが生
成する為生理的に有効なＬ一体の精製に複雑な工程を要
する。一方酵母菌体からの抽出法によるときは、所望の
Ｌ一体のみが得られる特徴がる。Glutathione has physiological effects such as liver function recovery and detoxification. It is also used in foods as a flavor enhancer. [Prior Art] As industrial methods for producing glutathione, a synthesis method and an extraction method from ban yeast and beer yeast cells have been adopted. However, when using a synthetic method, DL-glutathione is produced, and therefore a complicated process is required to purify physiologically effective L-unit. On the other hand, when using an extraction method from yeast cells, only the desired L monomer can be obtained.
酵母菌体からの抽出法によるグルクチオン生成において
は、亜鉛イオン制限下でグルタチオン生成蓄積量が著し
く増加したことが知られていた（特公昭５２−８３９７
号）。しかし、工業的には、培地由来の亜鉛イオン含量
が一定でない為に当該イオンの制御が難しく、グルタチ
オンの安定的な製造法が困難であった。In the production of glucthione by the extraction method from yeast cells, it was known that the amount of accumulated glutathione produced increased significantly under the restriction of zinc ions (Japanese Patent Publication No. 52-8397).
issue). However, industrially, since the content of zinc ions derived from the medium is not constant, it is difficult to control the ions, and it has been difficult to stably produce glutathione.
本発明の目的は、グルタチオン生成培地中の亜鉛イオン
を制御することなく、このグルタチオンを発酵工業的に
安価かつ安定的に製造する方法に関するものである。The object of the present invention is to provide a method for producing glutathione inexpensively and stably by fermentation without controlling zinc ions in a glutathione-producing medium.
（課題を解決する為の手段〕
本発明者らは、亜鉛イオンの制限範囲を拡大すべくサ・
７カロミセス属に属する酵母の亜鉛耐性を有する変異株
及び突然変異により亜鉛耐性を有する変異株が培地や培
養条件による影響を大幅に改善し安定したグルタチオン
生成蓄積し得ることを見い出し、本発明を完成するに至
った。(Means for Solving the Problems) The present inventors have developed a sa...
We have completed the present invention by discovering that mutant strains of yeast belonging to the genus Calomyces and mutant strains that have zinc tolerance due to mutation can significantly improve the effects of medium and culture conditions and can stably produce and accumulate glutathione. I ended up doing it.
すなわち、本発明は、グルタチオンを生産する能力を有
するサノ力ロミセス属の酵母より亜鉛イオン耐性を獲得
した変異株を培養し、グルタチオンを生成蓄積せしめる
ことを特徴とするグルタチオン高含有酵母の製造方法、
及び上記方法で製造されたグルタチオン高含有酵母に関
するものである。That is, the present invention provides a method for producing yeast with high glutathione content, which comprises culturing a mutant strain of yeast of the genus Sanoromyces that has the ability to produce glutathione and allowing it to produce and accumulate glutathione.
and glutathione-rich yeast produced by the above method.
以下に本発明を更に詳細に説明する。まずサソ力ロミセ
ス・セレビシエＡＪ−４ ０ ０ ５ （ ＦＥＲＭＰ
−１８５９）を用いてこれを紫外線，エチルメタンスル
ホネートニト口ソグアニジン，Ｘ線等の突然変異剤によ
り変異させた酵母菌、または無変異の酵母菌を、ディフ
コ製バクトＹＭ寒天培地に亜鉛イオンとして２０〜１０
０ｐｐｍ含んだ培地で２６〜３０゜ｃ， ２日間培養
し、生育した酵母菌を分離する。この際に使用する亜鉛
化合物としては、例えば硫酸亜鉛，酢酸亜鉛，塩化亜鉛
，乳酸亜鉛，燐酸亜鉛，ステアリン酸亜鉛等が用いられ
る。The present invention will be explained in more detail below. First of all, Sasoromyces cerevisiae AJ-4 0 0 5 (FERMP
-1859) and mutated with mutagens such as ultraviolet rays, ethylmethanesulfonate, nitrosoguanidine, and X-rays, or unmutated yeast were placed on Difco's Bacto YM agar medium as zinc ions for 20 min. ~10
Culture in a medium containing 0 ppm at 26-30°C for 2 days, and isolate the grown yeast. Examples of the zinc compound used in this case include zinc sulfate, zinc acetate, zinc chloride, zinc lactate, zinc phosphate, and zinc stearate.
本発明において使用する菌株は、サソカロミセス属酵母
で亜鉛イオン耐性を有する変異株のうち、親株に比して
蓄量のグルタチオン生成蓄積することができる変異株で
あり、例えばサソカロミセス・セレビシエＡ Ｊ −１
４６５２ （ＦＥＲＭ Ｐ−１０６９２）などがある
。The strain used in the present invention is a mutant strain of yeast of the genus Sasocharomyces that is resistant to zinc ions and is capable of producing and accumulating a larger amount of glutathione than the parent strain, such as Sasocharomyces cerevisiae A J-1.
4652 (FERM P-10692).
上記の如くグルタチオン生産能を有する微生物について
、亜鉛イオン含有量を第１表の如く変化せしめた培地で
培養したときの酵母の生育量を第１表に例示する。Table 1 shows the growth rate of yeast when the microorganisms having glutathione-producing ability as described above are cultured in media with zinc ion contents varied as shown in Table 1.
第１表から明らかな如く、該菌Ａ Ｊ−１４６５２は亜
鉛イオンに対して耐性であることが判る。As is clear from Table 1, the bacteria A J-14652 is resistant to zinc ions.
第 １ 表
次に、前述したグルタチオン生産能を有する微生物につ
いて、亜鉛イオン含有量を第２表の如く変化せしめた培
地で培養したときの菌体抽出液のグルタチオン生成量を
第２表に例示する。第２表から明らかな如く、グルタチ
オンの生成量に関しても問題ないことが判る。Table 1 Next, Table 2 shows the amount of glutathione produced in the bacterial cell extract when the aforementioned microorganisms with glutathione-producing ability are cultured in media with varying zinc ion contents as shown in Table 2. . As is clear from Table 2, there was no problem with the amount of glutathione produced.
第２表
？験条件：グルコース３％、ＫＩ１２Ｐ０． ０．
１ ５％，ＭｇＳＯ４・７１１ｚ０ ０． ０ ５％
，尿素０．３％， ＣａＣｊ２ｚ２１１■００．０５
％，ビオチン５０γ／ｌ，綜合アミノ酸０．２％及びビ
タミン混液１．Ｏｍ６／ｄｆｆを含有する培地をｐＨ６
．０に調整しその培地２０ｍｊ！を５００ｍｊ！振盪フ
ラスコに分注した后この培地に上記各菌株を接種し、３
０℃にて３０時間振盪培養した。生育は、培養ブロスを
２６倍希釈し平間式光電比色計を用いて５６２ｎｍにお
ける吸光度（ＯＤＳ６２）で測定される。Table 2? Test conditions: glucose 3%, KI12P0. 0.
1 5%, MgSO4・711z0 0. 0 5%
, Urea 0.3%, CaCj2z211■00.05
%, biotin 50γ/l, integrated amino acids 0.2% and vitamin mixture 1. The medium containing Om6/dff was adjusted to pH 6.
．． 0 and the medium was 20mj! 500mj! After dispensing into shake flasks, each of the above bacterial strains was inoculated into this medium, and 3
Shaking culture was performed at 0°C for 30 hours. Growth is measured by diluting the culture broth 26 times and measuring absorbance at 562 nm (ODS62) using a Hirama photoelectric colorimeter.
実験条件：グルコ−ス３％、κＩ１２ＰＯ４ ０．
１ ５ ％，ＭｇＳＯ＜ ・７１ｈ８ ０． ０
５％，尿素０． ３％， ＣａＣｌ，２Ｈ２０ ０
． ０ ５％，ビオチン５０ｒ／ｌ，綜合アミノ酸０．
２％及びビタミン混液１．Ｏｍｌ／ｄ／を含有する培地
をｐＨ６．０に調整しこの培地２０ｍ７！を５００ｍｊ
２振盪フラスコに分注した后この培地に上記各菌株を接
種し、３０℃にて３０時間振盪培養した。菌体抽出液グ
ルタチオン生成量の測定は、液体クロマトグラフィーに
よる。Experimental conditions: glucose 3%, κI12PO4 0.
15%, MgSO<・71h8 0. 0
5%, urea 0. 3%, CaCl, 2H20 0
．． 0.5%, biotin 50r/l, synthetic amino acids 0.
2% and vitamin mixture 1. A medium containing Oml/d/ was adjusted to pH 6.0 and this medium was 20m7! 500mj
After dispensing into two shaking flasks, each of the above-mentioned bacterial strains was inoculated into this medium and cultured with shaking at 30°C for 30 hours. The amount of glutathione produced in the bacterial cell extract was measured by liquid chromatography.
本発明を実施するには、上記の如く、特別に亜鉛イオン
含量を限定した培地を用いることなく、グルタチオン生
産能を有する微生物を通常の炭素源，窒素源および微生
物の必要とする無機並びに有機栄養素を含有する培地を
用いればよい。To carry out the present invention, as described above, without using a medium with a specially limited zinc ion content, microorganisms capable of producing glutathione are used as a source of ordinary carbon sources, nitrogen sources, and inorganic and organic nutrients required by microorganisms. A medium containing the following may be used.
用いられる炭素源としては、グルコース，フラクトース
，澱粉加水分解物．糖密等の［質，グリセロール，エタ
ノール等のアルコール、酢酸．クエン酸，α−ケトグル
タール酸等の有機酸、また菌株によっては炭化水素を用
いてもよい。Carbon sources used include glucose, fructose, and starch hydrolyzate. Sugar content, alcohol such as glycerol, ethanol, acetic acid. Organic acids such as citric acid and α-ketoglutaric acid, and depending on the strain, hydrocarbons may also be used.
培養は通常ρ１｛４〜８、温度２５〜３５℃にて好気的
に培養する。得られたグルタチオン高含有酵母は、医薬
・食品・機能性食品等に使用しても良い。更にグルタチ
オンの単離は、必要に応じて行なうことができる。Culture is usually carried out aerobically at ρ1{4 to 8 and a temperature of 25 to 35°C. The obtained glutathione-rich yeast may be used for medicines, foods, functional foods, etc. Furthermore, isolation of glutathione can be performed as necessary.
グルタチオンの単離法は、常法を用いれば良い．例えば
、菌体を遠心分離により集め、菌体のグルタチオン熱水
抽出処理やクロマト処理等を用いて単離することができ
る。Glutathione can be isolated using conventional methods. For example, the bacterial cells can be collected by centrifugation and isolated by subjecting the bacterial cells to glutathione hot water extraction treatment, chromatography treatment, or the like.
以下、本発明を実施例により具体的に説明する。Hereinafter, the present invention will be specifically explained with reference to Examples.
グルコース３％、ＫＨｚＰＯ＜ ｏ． １ ５％、Ｍ
ｇＳＯ．７ｎｚｏ Ｏ． ０ ５％，尿素０． ３％
， Ｃａ（Ｊ’ ｚ ’ ２ｔｈＯＯ．０５％，ビ
オチン５０γ／１，綜合アミノ酸０．２％およびビタミ
ン混液１，ＯｍＡ／ｄｊ２よりなる種培地を調製し、Ｎ
ａＯＩＩにてｐｌ＋６．０に調整した後、５００ＩＩＩ
１振盪フラスコに５０Ｉｌｊ！宛分注した。Glucose 3%, KHzPO<o. 15%, M
gSO. 7nzo O. 0.5%, urea 0. 3%
, Ca(J' z ' 2thOO.05%, biotin 50γ/1, integrated amino acids 0.2% and vitamin mixture 1, OmA/dj2 was prepared.
After adjusting to pl+6.0 with aOII, 500III
50 Ilj in 1 shake flask! Dispense to address.
殺菌後、亜鉛イオン耐性を有しないＡ Ｊ −４００５
（ＦＥＲＭ Ｐ−１８５９）、亜鉛イオン耐性を獲得し
たＡＪ−１４６５２ （ＦＥＲＭ Ｐ−１０６９２）を
それぞれ接種し、３０℃にて２０時間振盪培養し、これ
を種菌液とした。この種菌液１５ｍｊ！を、グルコース
３％，ＫＩｂＰＯ４ｏ． １ ５％， ＭｇＳＯ．・
７１１２０ ０．０５％，ＣａＣ ｌ ｚ ・２ＨＪ
Ｏ．０３％， （ＮＨｔ）ｚｓＯ４ ０．５％，ビオ
チン２０γ／ｌ，綜合アミノ酸０．２％，ビタミン混液
０．５ ｍ１／ ｄ ｊｌ！， ＦｅＳ０４１１１ｚｏ
Ｏ．０ ０ １％， ＭｎＳＯ ・４１１２０４
０．００１％及び亜鉛イオン０． ２ ｐｐｍ又は８．
３ ｐｐｍからなる本培養培地３００ｍｌに接種し、
通気Ｎ Ｉ／４ Ｖ／Ｖ，ｍ ，撹拌数１ ０ ０
０ｒｐｏ＋，３１℃にて２４時間培養した。培養中、
アンモニアガスを通してｐｌｌを５．０に制御した。こ
のときのグルタチオン生成量，菌体量等の結果を第３表
，第４表，第５表に示した。A J-4005 that does not have zinc ion resistance after sterilization
(FERM P-1859) and AJ-14652 (FERM P-10692) which had acquired zinc ion resistance were inoculated and cultured with shaking at 30° C. for 20 hours, and this was used as an inoculum solution. This seed liquid is 15mj! , glucose 3%, KIbPO4o. 15%, MgSO.・
71120 0.05%, CaClz・2HJ
O. 03%, (NHt)zsO4 0.5%, biotin 20γ/l, integrated amino acids 0.2%, vitamin mixture 0.5 m1/d jl! , FeS04111zo
O. 0 0 1%, MnSO ・411204
0.001% and zinc ion 0. 2 ppm or 8.
Inoculate 300 ml of main culture medium consisting of 3 ppm,
Aeration N I/4 V/V, m, stirring number 1 0 0
Cultured at 0rpo+, 31°C for 24 hours. During cultivation,
Pll was controlled at 5.0 by passing ammonia gas. The results of the glutathione production amount, bacterial cell amount, etc. at this time are shown in Tables 3, 4, and 5.
第４表
第３表
第５表
〔発明の効果〕
本発明の亜鉛イオン耐性を有するサッヵロミセス属酵母
を用いれば、培地等に由来する亜鉛イオン含量の変動に
左右されることもなく安定的にグルタチオン高含有酵母
または著量のグルタチオンを取得できることが明らかで
ある。Table 4 Table 3 Table 5 [Effects of the Invention] By using the yeast of the genus Saccharomyces that is resistant to zinc ions of the present invention, glutathione can be stably produced without being affected by fluctuations in the zinc ion content derived from the medium etc. It is clear that high yeast content or significant amounts of glutathione can be obtained.
Claims (1)
ス属の酵母より亜鉛イオン耐性を獲得した変異株を培養
し、グルタチオンを生成蓄積せしめることを特徴とする
グルタチオン高含有酵母の製造方法。 ２）請求項１記載の方法により製造されたグルタチオン
高含有酵母。[Scope of Claims] 1) A method for producing yeast with a high glutathione content, which comprises culturing a mutant strain of yeast of the genus Saccharomyces that has the ability to produce glutathione and which has acquired resistance to zinc ions, thereby producing and accumulating glutathione. 2) A glutathione-rich yeast produced by the method according to claim 1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP11529589A JP2830042B2 (en) | 1989-05-09 | 1989-05-09 | Glutathione-rich yeast and method for producing the same |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP11529589A JP2830042B2 (en) | 1989-05-09 | 1989-05-09 | Glutathione-rich yeast and method for producing the same |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPH02295480A true JPH02295480A (en) | 1990-12-06 |
| JP2830042B2 JP2830042B2 (en) | 1998-12-02 |
Family
ID=14659112
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP11529589A Expired - Lifetime JP2830042B2 (en) | 1989-05-09 | 1989-05-09 | Glutathione-rich yeast and method for producing the same |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JP2830042B2 (en) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE4219381A1 (en) * | 1992-06-13 | 1993-12-16 | Huels Chemische Werke Ag | High yield fermentative prodn. of glutathione - using zinc resistant mutants of Saccharomyces, specifically the new strain DSM 7092 |
| WO2004003217A1 (en) * | 2002-06-28 | 2004-01-08 | Unisearch Limited | Glutathione production |
| JP2008099578A (en) * | 2006-10-18 | 2008-05-01 | Medience Corp | Zinc yeast and method for producing the same |
| WO2010116833A1 (en) | 2009-04-08 | 2010-10-14 | Ajinomoto Co., Inc. | Novel yeast having increased content of sulfur-containing compound, screening method thereof, and culturing method thereof |
| WO2011129462A2 (en) | 2010-04-12 | 2011-10-20 | Ajinomoto Co., Inc. | A YEAST EXTRACT CONTAINING γ-Glu-X OR γ-Glu-X-Gly AND A METHOD FOR PRODUCING THE SAME |
| JP2014103855A (en) * | 2012-11-22 | 2014-06-09 | Medience Corp | Mineral yeast and production method thereof |
-
1989
- 1989-05-09 JP JP11529589A patent/JP2830042B2/en not_active Expired - Lifetime
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE4219381A1 (en) * | 1992-06-13 | 1993-12-16 | Huels Chemische Werke Ag | High yield fermentative prodn. of glutathione - using zinc resistant mutants of Saccharomyces, specifically the new strain DSM 7092 |
| WO2004003217A1 (en) * | 2002-06-28 | 2004-01-08 | Unisearch Limited | Glutathione production |
| JP2008099578A (en) * | 2006-10-18 | 2008-05-01 | Medience Corp | Zinc yeast and method for producing the same |
| WO2010116833A1 (en) | 2009-04-08 | 2010-10-14 | Ajinomoto Co., Inc. | Novel yeast having increased content of sulfur-containing compound, screening method thereof, and culturing method thereof |
| WO2011129462A2 (en) | 2010-04-12 | 2011-10-20 | Ajinomoto Co., Inc. | A YEAST EXTRACT CONTAINING γ-Glu-X OR γ-Glu-X-Gly AND A METHOD FOR PRODUCING THE SAME |
| US9034403B2 (en) | 2010-04-12 | 2015-05-19 | Ajinomoto Co., Inc. | Yeast extract containing γ-Glu-X or γ-Glu-X-Gly and a method for producing the same |
| JP2014103855A (en) * | 2012-11-22 | 2014-06-09 | Medience Corp | Mineral yeast and production method thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| JP2830042B2 (en) | 1998-12-02 |
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