JPS60156379A - Preparation of yeast having high gluthathione content - Google Patents
Preparation of yeast having high gluthathione contentInfo
- Publication number
- JPS60156379A JPS60156379A JP1200784A JP1200784A JPS60156379A JP S60156379 A JPS60156379 A JP S60156379A JP 1200784 A JP1200784 A JP 1200784A JP 1200784 A JP1200784 A JP 1200784A JP S60156379 A JPS60156379 A JP S60156379A
- Authority
- JP
- Japan
- Prior art keywords
- glutathione
- yeast
- gluthathione
- strain
- temperature
- 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
- 240000004808 Saccharomyces cerevisiae Species 0.000 title claims abstract description 20
- 241000222120 Candida <Saccharomycetales> Species 0.000 claims abstract description 16
- 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 claims description 84
- 108010024636 Glutathione Proteins 0.000 claims description 42
- 229960003180 glutathione Drugs 0.000 claims description 42
- 230000001580 bacterial effect Effects 0.000 claims description 13
- 238000012258 culturing Methods 0.000 claims description 8
- 238000004519 manufacturing process Methods 0.000 claims description 8
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 abstract description 6
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 abstract description 6
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 abstract description 6
- 238000009825 accumulation Methods 0.000 abstract description 5
- 241000235646 Cyberlindnera jadinii Species 0.000 abstract description 4
- 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 abstract description 4
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 abstract description 4
- 239000008103 glucose Substances 0.000 abstract description 4
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 abstract description 3
- 229910052943 magnesium sulfate Inorganic materials 0.000 abstract description 3
- 235000019341 magnesium sulphate Nutrition 0.000 abstract description 3
- 239000011591 potassium Substances 0.000 abstract description 3
- 229910052700 potassium Inorganic materials 0.000 abstract description 3
- 239000001103 potassium chloride Substances 0.000 abstract description 3
- 235000011164 potassium chloride Nutrition 0.000 abstract description 3
- 229910000147 aluminium phosphate Inorganic materials 0.000 abstract description 2
- 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 abstract description 2
- 229910002651 NO3 Inorganic materials 0.000 abstract 1
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 abstract 1
- YYRMJZQKEFZXMX-UHFFFAOYSA-L calcium bis(dihydrogenphosphate) Chemical compound [Ca+2].OP(O)([O-])=O.OP(O)([O-])=O YYRMJZQKEFZXMX-UHFFFAOYSA-L 0.000 abstract 1
- 229910000389 calcium phosphate Inorganic materials 0.000 abstract 1
- 229910017053 inorganic salt Inorganic materials 0.000 abstract 1
- 235000019691 monocalcium phosphate Nutrition 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 19
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 4
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- LSNNMFCWUKXFEE-UHFFFAOYSA-N Sulfurous acid Chemical compound OS(O)=O LSNNMFCWUKXFEE-UHFFFAOYSA-N 0.000 description 4
- 238000000034 method Methods 0.000 description 4
- 230000000813 microbial effect Effects 0.000 description 4
- LWIHDJKSTIGBAC-UHFFFAOYSA-K tripotassium phosphate Chemical compound [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- GGLZPLKKBSSKCX-YFKPBYRVSA-N L-ethionine Chemical compound CCSCC[C@H](N)C(O)=O GGLZPLKKBSSKCX-YFKPBYRVSA-N 0.000 description 3
- 229940041514 candida albicans extract Drugs 0.000 description 3
- 230000000052 comparative effect Effects 0.000 description 3
- 239000000284 extract Substances 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 239000012138 yeast extract Substances 0.000 description 3
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 description 2
- 238000005273 aeration Methods 0.000 description 2
- 229910021529 ammonia Inorganic materials 0.000 description 2
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 2
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 2
- 235000011130 ammonium sulphate Nutrition 0.000 description 2
- -1 and the like Substances 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 239000013078 crystal Substances 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 230000035772 mutation Effects 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 229910000160 potassium phosphate Inorganic materials 0.000 description 2
- 235000011009 potassium phosphates Nutrition 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 210000005253 yeast cell Anatomy 0.000 description 2
- 229910052725 zinc Inorganic materials 0.000 description 2
- 239000011701 zinc Substances 0.000 description 2
- MIQITCBUROTPKR-GEMLJDPKSA-N (2s)-2-amino-5-[[(2r)-1-(carboxymethylamino)-1-oxo-3-sulfanylpropan-2-yl]amino]-5-oxopentanoic acid;copper Chemical compound [Cu].OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O MIQITCBUROTPKR-GEMLJDPKSA-N 0.000 description 1
- ZCYVEMRRCGMTRW-UHFFFAOYSA-N 7553-56-2 Chemical compound [I] ZCYVEMRRCGMTRW-UHFFFAOYSA-N 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 235000008733 Citrus aurantifolia Nutrition 0.000 description 1
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 1
- RWSOTUBLDIXVET-UHFFFAOYSA-N Dihydrogen sulfide Chemical compound S RWSOTUBLDIXVET-UHFFFAOYSA-N 0.000 description 1
- 229930189936 Glyoxalase Natural products 0.000 description 1
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 description 1
- PWHULOQIROXLJO-UHFFFAOYSA-N Manganese Chemical compound [Mn] PWHULOQIROXLJO-UHFFFAOYSA-N 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- 235000011941 Tilia x europaea Nutrition 0.000 description 1
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- LFVGISIMTYGQHF-UHFFFAOYSA-N ammonium dihydrogen phosphate Chemical compound [NH4+].OP(O)([O-])=O LFVGISIMTYGQHF-UHFFFAOYSA-N 0.000 description 1
- QLULGSLAHXLKSR-UHFFFAOYSA-N azane;phosphane Chemical compound N.P QLULGSLAHXLKSR-UHFFFAOYSA-N 0.000 description 1
- YYRMJZQKEFZXMX-UHFFFAOYSA-N calcium;phosphoric acid Chemical compound [Ca+2].OP(O)(O)=O.OP(O)(O)=O YYRMJZQKEFZXMX-UHFFFAOYSA-N 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 239000003518 caustics Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 229910052802 copper Inorganic materials 0.000 description 1
- 239000010949 copper Substances 0.000 description 1
- 150000001879 copper Chemical class 0.000 description 1
- 229910000365 copper sulfate Inorganic materials 0.000 description 1
- BERDEBHAJNAUOM-UHFFFAOYSA-N copper(I) oxide Inorganic materials [Cu]O[Cu] BERDEBHAJNAUOM-UHFFFAOYSA-N 0.000 description 1
- ARUVKPQLZAKDPS-UHFFFAOYSA-L copper(II) sulfate Chemical compound [Cu+2].[O-][S+2]([O-])([O-])[O-] ARUVKPQLZAKDPS-UHFFFAOYSA-L 0.000 description 1
- OMZSGWSJDCOLKM-UHFFFAOYSA-N copper(II) sulfide Chemical compound [S-2].[Cu+2] OMZSGWSJDCOLKM-UHFFFAOYSA-N 0.000 description 1
- KRFJLUBVMFXRPN-UHFFFAOYSA-N cuprous oxide Chemical compound [O-2].[Cu+].[Cu+] KRFJLUBVMFXRPN-UHFFFAOYSA-N 0.000 description 1
- 229940112669 cuprous oxide Drugs 0.000 description 1
- 238000001784 detoxification Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 239000011790 ferrous sulphate Substances 0.000 description 1
- 235000003891 ferrous sulphate Nutrition 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 229910000037 hydrogen sulfide Inorganic materials 0.000 description 1
- 239000011630 iodine Substances 0.000 description 1
- 229910052740 iodine Inorganic materials 0.000 description 1
- XEEYBQQBJWHFJM-UHFFFAOYSA-N iron Substances [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 description 1
- 229910000359 iron(II) sulfate Inorganic materials 0.000 description 1
- 239000004571 lime Substances 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 230000003908 liver function Effects 0.000 description 1
- 239000011777 magnesium Substances 0.000 description 1
- 229910052749 magnesium Inorganic materials 0.000 description 1
- 229910001629 magnesium chloride Inorganic materials 0.000 description 1
- 229910052748 manganese Inorganic materials 0.000 description 1
- 239000011572 manganese Substances 0.000 description 1
- 229940099596 manganese sulfate Drugs 0.000 description 1
- 239000011702 manganese sulphate Substances 0.000 description 1
- 235000007079 manganese sulphate Nutrition 0.000 description 1
- SQQMAOCOWKFBNP-UHFFFAOYSA-L manganese(II) sulfate Chemical compound [Mn+2].[O-]S([O-])(=O)=O SQQMAOCOWKFBNP-UHFFFAOYSA-L 0.000 description 1
- 235000013379 molasses Nutrition 0.000 description 1
- 150000002823 nitrates Chemical class 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 229920001296 polysiloxane Polymers 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 239000002426 superphosphate Substances 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 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】
本発明は、グルタチオンを著量含有する酵母の製造法に
関するものである。DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method for producing yeast containing significant amounts of glutathione.
更に詳細には、本発明は、キャンデイダ属に属するグル
タチオン生産性酵母を該菌株の生育至適温度よシ低い培
養温度にて好気的に培養することにより、該菌体中に著
量のグルタチオンを生成蓄積させた酵母を製造する方法
に関するものである。More specifically, the present invention cultivates a glutathione-producing yeast belonging to the genus Candida aerobically at a culture temperature lower than the optimum growth temperature of the strain, thereby cultivating a significant amount of glutathione in the bacterial cells. This invention relates to a method for producing yeast that produces and accumulates .
一般にグルタチオンは酵母及び動物の肝臓などに広く分
布しており、生体内の酸化還元系に関与しているトリは
プチドで、肝機能回復作用や解毒作用などの重要表役割
を果す医薬上極めて有用な物質である。In general, glutathione is widely distributed in yeast and animal livers, and is involved in the redox system in living organisms, and is extremely useful medicinally as it plays an important role in restoring liver function and detoxification. It is a substance.
本発明の目的は、このグルタチオンを発酵工業的に有利
に製造する方法を提供することにある。An object of the present invention is to provide a method for producing glutathione advantageously in the fermentation industry.
近年、グルタチオンはほとんどが酵母菌体からの抽出方
法によシ生産されており、酵母菌体中のグルタチオン含
量を高める方法が数多く提案されているが、いずれも培
養における培養温度は酵母の至適生育温度である３０℃
前後にて実施されている。In recent years, most glutathione has been produced by extraction from yeast cells, and many methods have been proposed to increase the glutathione content in yeast cells. Growth temperature of 30℃
It is carried out before and after.
本発明者らは、キャンデイダ属におけるグルタチオンの
蓄積量を高めるために鋭意研究を行った結果、酵母を培
養する際、該菌株の生育至適温度よシ低い培養温度で培
養することによりグルタチオンの蓄積量を著しく高める
ことができることを知ったのである。The present inventors have conducted extensive research to increase the amount of glutathione accumulated in Candida spp., and have found that when culturing yeast, glutathione is accumulated by culturing at a culture temperature lower than the optimum growth temperature of the strain. I learned that I could significantly increase the amount.
更に研究を進めたところ、酵母菌株の培養適温よシかな
り低いところにグルタチオン蓄積適温があることも知っ
たのである。Further research led them to discover that the optimum temperature for glutathione accumulation is considerably lower than the optimum culture temperature for yeast strains.
本発明は、これら知見から完成されたもので、キャンデ
イダ属に属するグルタチオン生産性酵母を該菌株の生育
適温度より低い培養温度で培養することにより、該菌体
中に著量の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法である。The present invention was completed based on these findings, and by culturing glutathione-producing yeast belonging to the genus Candida at a culture temperature lower than the optimum growth temperature of the strain, a significant amount of glutathione is produced and accumulated in the bacterial body. This is a method for producing yeast with high glutathione content.
そして、本発明では、キャンデイダ属に属するグルタチ
オン生産性酵母がエチオニンおよび亜硫酸塩を含む培地
に生育可能となった突然変異株の使用が好ましい。In the present invention, it is preferable to use a mutant strain of glutathione-producing yeast belonging to the genus Candida that can grow in a medium containing ethionine and sulfite.
また、本発明では培養温度が至適生育温度より５℃以上
低い培養温度であることが好ましく、具体的には、培養
温度が１８〜２５℃であることが好ましい。Further, in the present invention, the culture temperature is preferably 5°C or more lower than the optimum growth temperature, and specifically, it is preferable that the culture temperature is 18 to 25°C.
本発明に使用するキャンデイダ属菌株はグルタチオンを
生産蓄積する菌株であればどのような菌株でもよいが、
突然変異処理によってエチオニンおよび亜硫酸塩を含む
培地に生育可能となった菌株（特願昭５８−２４８１５
）は特にグルタチオンを著量蓄積できるので好ましい。The Candida strain used in the present invention may be any strain as long as it produces and accumulates glutathione.
A strain that can grow in a medium containing ethionine and sulfite through mutation treatment (Patent application No. 58-24815)
) is particularly preferred because it can accumulate a significant amount of glutathione.
突然変異処理によってエチオニンおよび亜硫酸塩を含む
培地に生育可能となった菌株としては、例えばキャンデ
イク・ウチルスＫＪ８−０５７１やキャンデイダ・ウチ
ルスＫ Ｊ Ｓ−０５８２などが有利に使用される。な
おキャンデイダ・ウチルスＫＪＳ−０５７１およびキャ
ンデイダ・ウチルスＫＪＳ−０５，８２は工業技術院微
生物工業技術研究所にＦｌ、Ｍ Ｐ −６９０７および
’Ｆ’ＥＲ，Ｍ Ｐ−７３９６として寄託されている。As strains that can grow in a medium containing ethionine and sulfite through mutation treatment, for example, Candida uchilus KJ8-0571 and Candida uchilus KJ S-0582 are advantageously used. In addition, Candida uchilus KJS-0571 and Candida uchilus KJS-05,82 have been deposited with the Institute of Microbial Technology, Agency of Industrial Science and Technology as Fl, MP-6907 and 'F'ER, MP-7396.
培養に用いられる炭素源としては、ブドウ糖、蔗糖、酢
酸、エタノール、糖蜜、亜硫酸パルプ廃液等が用いられ
、また窒素源としては、アンモニア、硫安、尿素、硝酸
塩などが使用される。さらに無機塩としては、燐酸、カ
リウム、マグネシウム源として、過リン酸石灰、燐安、
塩化カリ、燐酸カリ、苛性カリ、硫酸マグネシウム、塩
化マグネシウム等が用いられ、さらに微量の亜鉛、銅、
マンガン、鉄イオン等の無機塩も使用される。ビタミン
、アミノ酸、核酸関連物質は特に必要としないが、勿呻
これらを添加したシコーンステイープリカー、酵母エキ
ス、ペプトン等を加えても差支えない。−は６．５〜８
０、特に４，０〜６．０が望ましい。Carbon sources used in the culture include glucose, sucrose, acetic acid, ethanol, molasses, sulfite pulp waste liquid, and the like, and nitrogen sources include ammonia, ammonium sulfate, urea, nitrates, and the like. Furthermore, as inorganic salts, as sources of phosphoric acid, potassium, and magnesium, lime superphosphate, ammonium phosphorus,
Potassium chloride, potassium phosphate, caustic potassium, magnesium sulfate, magnesium chloride, etc. are used, and trace amounts of zinc, copper,
Inorganic salts such as manganese and iron ions are also used. Vitamins, amino acids, and nucleic acid-related substances are not particularly required, but silicone staple liquor, yeast extract, peptone, etc. containing these may of course be added. - is 6.5 to 8
0, especially 4.0 to 6.0.
培養温度として、グルタチオン生産性菌株の生育至適温
度より低い温度であればグルタチオンはより多く蓄積さ
れるのである。一般的には菌株の至適生育温度より５℃
以上低い温度がグルタチオンの蓄積には適している。そ
して、グルタチオンの蓄積だけを考慮するならば培養温
度は１８〜２５℃が最も好ましいものである。If the culture temperature is lower than the optimal growth temperature of the glutathione-producing strain, more glutathione will be accumulated. Generally, 5℃ below the optimal growth temperature of the strain.
Temperatures as low as this are suitable for accumulating glutathione. If only the accumulation of glutathione is considered, the most preferable culture temperature is 18 to 25°C.
本発明における培養温度は菌株の至適生育温度よシ低い
ために、酵母の増殖はおそくなる傾向にある。従って、
培養温度を低くする場合は培養時間を長くする必要があ
る。菌株によってもそれぞれ異なるが、一般的には２５
〜５０時間程度である。Since the culture temperature in the present invention is lower than the optimum growth temperature of the bacterial strain, the growth of yeast tends to be slow. Therefore,
When lowering the culture temperature, it is necessary to lengthen the culture time. Although it varies depending on the strain, generally 25
~50 hours.
またこの際の培養形式は好気培養であればバッチ培養、
或は連続培養の倒れでもよい。In addition, the culture format at this time is batch culture if aerobic culture,
Alternatively, continuous culture may be collapsed.
次に、試験例、実施例及び比較例をあげて説明する。な
お、グルタチオンの定量は、菌体抽出液についてグリオ
キサラーゼ法（１−メソッド・イン。Next, test examples, examples, and comparative examples will be given and explained. Incidentally, glutathione was quantified using the glyoxalase method (1-Method In) for the bacterial cell extract.
エンザイモロジー」第１巻５４０に一ジ、アカデミツク
ブレス社、１９５５年版）で行った。Enzymology, Volume 1, 540, Academic Press, 1955 edition).
まだ本発明の有用性は、以下の実施例に示すが、これに
よって本発明が制限されるものではない。The utility of the present invention is further illustrated by the following examples, which do not limit the invention.
試験例１゜
キャンデイダ・ウチルスＫＪ８−０５８２株ｌＲ，Ｍ
Ｐ−７３９６を用い培養温度を１８℃〜３４℃の９段階
に変化させて、あとの条件は、スケールを１／１０とし
た以外は実施例１と同様に試験して各培養温度における
増殖速度及びグルタチオン含有量を測定した。増殖速度
は３２℃における対数増殖期の比増殖速度（０６９３／
ダブリングタイム）を１００％とし、各培養温度におけ
る比増殖速度を相対値としてめた。またグルタチオン含
量は各培養温度での培養後了後の菌体をそれぞれ集め、
実施例１と同様に処理してグルタチオン含量をめた。Test example 1 Candida uchilus KJ8-0582 strain 1R, M
Using P-7396, the culture temperature was varied in nine stages from 18°C to 34°C, and the remaining conditions were the same as in Example 1 except that the scale was 1/10, and the growth rate at each culture temperature was determined. and glutathione content was measured. The growth rate is the specific growth rate (0693/
Doubling time) was set as 100%, and the specific growth rate at each culture temperature was determined as a relative value. In addition, the glutathione content was determined by collecting the bacterial cells after culturing at each culture temperature.
The glutathione content was determined in the same manner as in Example 1.
その結果は第１図に示される通ゆであるが、ＫＪ Ｓ
−０５８２株の場合、生育至適温度が６２℃であシ、グ
ルタチオン蓄積の至適温度は２２９Ｃ〜２４℃であるこ
とがわかる。The results are shown in Figure 1, but KJ S
In the case of strain -0582, the optimum temperature for growth is 62°C, and the optimum temperature for glutathione accumulation is 229°C to 24°C.
試験例２゜
キャンデイダ・ウチリスＫＪ Ｓ−０５７１株、ＦＥＢ
Ｍ Ｐ−６９０７を用い、試験例１と同様にして相対比
増殖速度（係）とグルタチオン含有量をめた。Test Example 2 Candida utilis KJ S-0571 strain, FEB
Using M P-6907, the relative specific growth rate (correspondence) and glutathione content were determined in the same manner as in Test Example 1.
その結果は第２図に示される通りであるが、ＫＪ Ｓ
−０５７１株の場合、生育至適温度が３０〜３２℃で、
グルタチオン蓄積の至適温度は２２℃であることがわか
る。The results are shown in Figure 2, and KJ S
In the case of -0571 strain, the optimum growth temperature is 30-32℃,
It can be seen that the optimum temperature for glutathione accumulation is 22°C.
実施例１゜
キャンデイダ・ウチルスＫＪＳ−０５８２株、ＦＥＢＭ
Ｐ−’ｒ：３Ｑ６ をグルコース２チイーストエキス
２％はプトン１チからなる培地でフラスコ培養し、これ
を３００を容発酵槽に１％植菌した。Example 1 Candida uchilus KJS-0582 strain, FEBM
P-'r:3Q6 was cultured in a flask in a medium consisting of 2% glucose, 2% yeast extract, and 1% yeast extract, and 1% of this was inoculated into a 300-volume fermenter.
培地としては、グルコース３係、硫安０，８チ、リン酸
−カリウム０．２％、硫酸マグネシウム０．０３係、硫
酸第一鉄１０ｐｐｍ、髄酸亜鉛３ ｐｐｍ、硫酸銅ｌｐ
ｐｍ、硫酸マンガン１０ｐｐｍを用いパッチ培養を行っ
た。培養条件としては槽内液量２００ ｔ。The medium contains glucose 3 parts, ammonium sulfate 0.8 parts, potassium phosphate 0.2%, magnesium sulfate 0.03 parts, ferrous sulfate 10 ppm, zinc myelate 3 ppm, copper sulfate lp.
Patch culture was performed using 10 ppm of manganese sulfate. The culture conditions were a tank liquid volume of 200 t.
培養温度２４℃、通気量１５０ｔｐｍ、撹拌数２００ｒ
ｐｍ、ｐＨ４，５にて行った。２８時間後集菌したとこ
ろ乾燥時換算２９４Ｏｆの菌体が得られ、菌体中のグル
タチオン含量は５．０％（対乾燥菌体比）であった。こ
の菌体を加熱抽出し、菌体残渣を遠心分離にて除きグル
タチオン抽出液を得た。この抽出液に硫酸を０．５ Ｎ
になるように添加し、亜酸化銅を加えグルタチオンを銅
塩として析出させた。Culture temperature 24℃, aeration rate 150tpm, stirring number 200r
pm and pH 4 and 5. When the bacteria were collected after 28 hours, 294 Of cells were obtained in dry terms, and the glutathione content in the cells was 5.0% (ratio to dry cells). The cells were heated and extracted, and the cell residue was removed by centrifugation to obtain a glutathione extract. Add 0.5 N of sulfuric acid to this extract.
cuprous oxide was added to precipitate glutathione as a copper salt.
グルタチオン銅塩を水洗した後硫化水素を通気し、硫化
銅を除き、減圧濃縮することによシグルタチオンの結晶
１２０ｆを得た。After washing the glutathione copper salt with water, hydrogen sulfide was bubbled through to remove the copper sulfide, and the mixture was concentrated under reduced pressure to obtain siglutathione crystals 120f.
得られた結晶は高圧ろ紙電気泳動、高速液体クロマトグ
ラフィーよシグルタチオンであることが確認され、ヨー
ド法による純度は９９０％であった。The obtained crystals were confirmed to be siglutathione by high pressure filter paper electrophoresis and high performance liquid chromatography, and the purity by the iodine method was 990%.
比較例１゜
キャンデイダ・ウチルスＫＪＳ−０５８２株、ＦＥＲＭ
Ｐ−’ｒ３９６ 株を培養温度６０℃以外は実施例１
と全く同様に培養し比較培養した。２２時間培養後２９
５０ｔの菌体（乾燥時換算）を得た。Comparative Example 1 Candida uchilus KJS-0582 strain, FERM
Example 1 except for culturing the P-'r396 strain at a temperature of 60°C.
The cells were cultured in exactly the same manner as above for comparison. After 22 hours of culture 29
50 tons of bacterial cells (calculated when dry) were obtained.
この菌体中のグルタチオン含量は６．７％（対乾燥菌体
）であった。実施例１と全く同様の処理により８７１の
グルタチオンを得た。The glutathione content in the bacterial cells was 6.7% (based on dry bacterial cells). Glutathione 871 was obtained by the same treatment as in Example 1.
実施例２
キャンデイダ・ウチルスＫＪＳ−０５７１株、Ｐ’ＷＲ
Ｍ Ｐ”６９０７を実施例１と同様にフラスコ種母培養
し３ｏｔ発酵槽に５１植菌した。Example 2 Candida uchilus KJS-0571 strain, P'WR
M P"6907 was cultured in a flask in the same manner as in Example 1, and 51 inoculated into a 3 ot fermenter.
培地としては次の組成のものを用いた。亜硫酸パルプ廃
液（資化性糖として６チ）にリン酸−アンモニウム０．
１５％、塩化カリウム０．０６％を添加する。培養はド
ラフトチューブ付発酵槽で槽内液−１ｉｏｚ、培養温度
２２℃通気量ｉＱｔｐｍ、撹拌数７０ Ｏｒｐｍで行い
、アンモニアを添加して…コントロール及び、培養の窒
素源とした。培養４２時間後に遠心分離にて菌体を集菌
したところ菌体１６０ｆが得られ菌体中のグルタチオン
含量は４．６％（対乾燥菌体比）であった。A medium with the following composition was used. Phosphoric acid-ammonium 0.0.
15% and potassium chloride 0.06%. The culture was carried out in a fermenter with a draft tube at -1 ioz of tank liquid, culture temperature of 22°C, aeration rate of iQtpm, and stirring number of 70 Orpm, and ammonia was added to serve as a control and a nitrogen source for culture. After 42 hours of culturing, the bacterial cells were collected by centrifugation to obtain 160 f bacterial cells, and the glutathione content in the bacterial cells was 4.6% (ratio to dry bacterial cells).
比較例２゜
キャンデイダ・ウチルスＫＪ Ｓ −０５７１株を培養
温度３０℃以外は実施例２と全く同様に培養し比較培養
した。５２時間培養後１６５ｆの菌体（乾燥時換算）が
得られ、菌体中のグルタチオン含量は３．８％（対乾燥
菌体比）であった。Comparative Example 2 Candida uchilus KJ S-0571 strain was cultured in the same manner as in Example 2 except that the culture temperature was 30°C for comparison. After culturing for 52 hours, 165 f microbial cells (calculated on dry basis) were obtained, and the glutathione content in the microbial cells was 3.8% (ratio to dry microbial cells).
第１図は試験例１においてキャンデイダ・ウチリスＫＪ
Ｓ−［１５８２株の相対比増殖速度（チ）とグルタチオ
ン含量ｆ％）をめた図で、第２図は試験例２においてキ
ャンデイダ・ウチリスＫＪ Ｓ −０５７１株の相対比
増殖速度部）とグルタチオン含量（チ）をめた図である
。
Ａ・・・相対比増殖速度部）
Ｂ・・・グルタチオン含量（％）
代理人 弁理士 戸 １）親 男
第 １ 図
培養温度（℃）
第 ２ 図
培養温度（℃）Figure 1 shows Candida utilis KJ in Test Example 1.
Figure 2 shows the relative specific growth rate (chi) and glutathione content f% of the Candida utilis strain KJ S-0571 in Test Example 2. It is a diagram showing the content (chi). A...Relative growth rate) B...Glutathione content (%) Agent Patent attorney 1) Parent Male Figure 1 Culture temperature (℃) Figure 2 Culture temperature (℃)
Claims (1)
を該菌株の生育至適温度よシ低い培養温度で培養するこ
とにより、該菌体中に著量の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
方九（２）キャンデイダ属に属するグルタチオン生産性
酵母がエチオニンおよび亜硫酸塩を含む培地に生育可能
となった突然変異株である特許請求の範囲第１項記載の
グルタチオン高含有酵母の製造法。 （３）培養温度が至適生育温度よシ５℃以上低い培養温
度であることを特徴とする特許請求の範囲第１項記載の
グルタチオン高含有酵母の製造法。 （４）培養温度が１８〜２５℃であることを特徴とする
特許請求の範囲第１項記載のグルタチオン高含有酵母の
製造法。[Scope of Claims] (1) By culturing glutathione-producing yeast belonging to the genus Candida at a culture temperature lower than the optimum growth temperature of the strain, a significant amount of glutathione is produced and accumulated in the bacterial cells. Method for producing yeast with high glutathione content, characterized by A method for producing yeast with high glutathione content. (3) The method for producing glutathione-rich yeast according to claim 1, wherein the culture temperature is 5° C. or more lower than the optimum growth temperature. (4) The method for producing glutathione-rich yeast according to claim 1, wherein the culture temperature is 18 to 25°C.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP1200784A JPS60156379A (en) | 1984-01-27 | 1984-01-27 | Preparation of yeast having high gluthathione content |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP1200784A JPS60156379A (en) | 1984-01-27 | 1984-01-27 | Preparation of yeast having high gluthathione content |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS60156379A true JPS60156379A (en) | 1985-08-16 |
| JPH0318872B2 JPH0318872B2 (en) | 1991-03-13 |
Family
ID=11793527
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP1200784A Granted JPS60156379A (en) | 1984-01-27 | 1984-01-27 | Preparation of yeast having high gluthathione content |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS60156379A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS6434279A (en) * | 1987-07-29 | 1989-02-03 | Ajinomoto Kk | Production of yeast with high glutathione content |
| JPH01141591A (en) * | 1987-11-26 | 1989-06-02 | Ajinomoto Co Inc | Production of yeast with high glutathione content |
| WO2019181961A1 (en) | 2018-03-20 | 2019-09-26 | 三菱商事ライフサイエンス株式会社 | METHOD FOR PRODUCING β-NMN AND COMPOSITION CONTAINING SAME |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| MY142328A (en) | 2002-03-26 | 2010-11-15 | Ajinomoto Kk | CANDIDA UTILIS CONTAINING y-GLUTAMYLCYSTEINE |
-
1984
- 1984-01-27 JP JP1200784A patent/JPS60156379A/en active Granted
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS6434279A (en) * | 1987-07-29 | 1989-02-03 | Ajinomoto Kk | Production of yeast with high glutathione content |
| JPH01141591A (en) * | 1987-11-26 | 1989-06-02 | Ajinomoto Co Inc | Production of yeast with high glutathione content |
| WO2019181961A1 (en) | 2018-03-20 | 2019-09-26 | 三菱商事ライフサイエンス株式会社 | METHOD FOR PRODUCING β-NMN AND COMPOSITION CONTAINING SAME |
Also Published As
| Publication number | Publication date |
|---|---|
| JPH0318872B2 (en) | 1991-03-13 |
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