JPS5836397A - Preparation of s-adenosyl methionine - Google Patents
Preparation of s-adenosyl methionineInfo
- Publication number
- JPS5836397A JPS5836397A JP13456181A JP13456181A JPS5836397A JP S5836397 A JPS5836397 A JP S5836397A JP 13456181 A JP13456181 A JP 13456181A JP 13456181 A JP13456181 A JP 13456181A JP S5836397 A JPS5836397 A JP S5836397A
- Authority
- JP
- Japan
- Prior art keywords
- methionine
- alanine
- glycine
- yeast
- culture medium
- 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
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
Description
【発明の詳細な説明】
本発明は醗酵法による8−アデノフルメチオニン(以下
、RAMと略称する)の製造方法に関し、更に詳しくは
、グリシン及び/またはアラニン系の低級ペプチドを添
加したメチオニ/含有液体培地中で酵母を培養し、8ム
Mt効率的に製造する方法に関する。DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method for producing 8-adenoflumethionine (hereinafter referred to as RAM) by a fermentation method, and more specifically, it relates to a method for producing 8-adenoflumethionine (hereinafter referred to as RAM) by a fermentation method. The present invention relates to a method of culturing yeast in a liquid medium to efficiently produce 8 μM Mt.
111Mは生体内において脂肪、蛋白質、種類などの代
−に関与する重要な物質である。面して近時かかる8ム
MK肝血痰、過度脂血症、動脈硬化症、抑うつ病訃よび
神経病形の精神病発現、変性閾接症神経病麿覚、不眠症
などに対する治療効果ohることが見い出されてお9、
その大量生産が期待されている。111M is an important substance involved in the production of fat, protein, and other substances in living organisms. Recently, it has been found to be effective in treating 8 mu mk liver hemoptysis, hyperlipidemia, arteriosclerosis, depressive illness, neurological manifestations of psychosis, degenerative threshold syndrome, neurological disorders, insomnia, etc. has been found9,
Mass production is expected.
従来、81M01111方法としては種々O黴生物を用
−る培養法が知られているが、なかでも酵母を用いる方
法が好ましいとされて訃)、その具体例としてナツカa
wイセス属ま九紘キャンディf属O黴生物を用いる方法
(91えばJowr+a亀1of11a*tari@1
6gy、マ・z 121a 247頁(1975年)な
ど)、ビヒア属、諺ドトルツ属、クリプトコツカス属、
ハ/ゼヌツ属、トリコスポロン属、フレケラ属、ハンゼ
ニアスポツ属、スポロSatセス属、リボ(セス属i九
紘デバリオを竜ス属の微生物を用いる方法(41会l!
1i52−17118号)などが知られているoしかし
ながら、これらO方法においても8AMの蓄積量は必ず
しも充分満足できるものとはいえず、より一層の改良が
求められていた。Conventionally, as the 81M01111 method, various culturing methods using fungal organisms have been known, but among them, the method using yeast is considered to be preferable.
w A method using genus Ises, Candy f, genus O fungi (91, for example, Jowr+a turtle1of11a*tari@1
6gy, Ma.z 121a, p. 247 (1975), etc.), genus Vichia, genus Dotorz, genus Cryptococcus,
A method using microorganisms of the genus Ha/Trichosporon, Frechella, Hanseniaspotus, SporoSat, and Ribo (41 meetings)
1i52-17118), etc. However, even in these O methods, the amount of 8AM accumulated is not necessarily fully satisfactory, and further improvements have been sought.
そこで本発明者らはmil法によるSAMの蓄積量向上
につき鋭意検討を加えた結果、SAMの前駆体であるメ
チオニ/とともKある種の低級ペプチドを添加した培地
を用いることが有効であることを見い出し、本発明を完
成し九。Therefore, the present inventors conducted extensive studies on improving the amount of SAM accumulated using the mil method, and found that it is effective to use a medium supplemented with a certain type of lower peptide, methionine/tomoK, which is a precursor of SAM. discovered this and completed the present invention.
すなわち本発明の目的は生産能に優れた発酵法による8
AMの製造方法を提供することにあり、かかる本発明の
目的は、8ムM生産能を有する酵母をメチオニン含有培
地で培養して13AMを製造する方法に>tnて、グリ
シ/及び/lたはアラニ/から構成される低級ペプチド
を培地に添加することによ)達成される。In other words, the purpose of the present invention is to produce 8
An object of the present invention is to provide a method for producing AM, and an object of the present invention is to provide a method for producing 13AM by culturing yeast capable of producing 8muM in a methionine-containing medium. is achieved by adding to the medium a lower peptide consisting of Arani/.
本発明において用いられる酵母はメチオニン含有培地中
でflAMを蓄積する能力を有するものであればいずれ
でもよいが、なかでもサツカロマイに属する酵母が好ま
しく、その具体例としてナッカロマイセス争セレビジエ
エpo 2546、キャンディダΦ!セドニエ/シス
IFO0960,ハ/ゼヌツ・ファビアニイIP0 1
570 などが′挙げられる。tlI−これらの天然
及び人工変異菌であってもBAM生書能を有するかぎシ
同様に使用することがで龜る・
本発明において紘、メチオニン含有培地中にグリシン単
独、アラ二ノ単独ま九はグリシン及びアラニンから構成
される低級ペプチドを添加することが必SOW件である
・ここでいう低級ペプチドとは、ジペプチド、トリペプ
チド及びテトラペプチドをさし、こOような低級ペプチ
ドの具体例として、例えばグリシルグリシン、グリクル
グリシルグリシン、グリVルダリシルグリシルグリシン
、D、L−アツエルD、L−アク二ノ、L−アツニルL
−アフニルL−72二ン、L−アラニルL−アラニルL
−アラニルL−79二ン、L−72ニルグリシ/などが
挙げられる・これらO低級ペプチドは通當培地中K(L
O2#/jJ以上の濃度で添加され、とくにcLo41
/dl 以上の濃度とするのが適切である・
本発明で用いる培地の他の条件は常法に従えばよ七、例
えば炭素源としてグルコース、シュクロース、7ラクト
ース等の糖類、酢酸などの有機酸類、炭化水素類、メタ
ノール、エタノールなどのアルコール類などを用いるこ
とができる。また培地中には更に通常の窒素源、無機塩
、有機微量栄養素が必要に応じて使用されるO
培養は好気的条件が良く、培養温度は20[11’から
40Cの範囲が好ましい。培養の際、培地のPHを3か
ら8の範囲に調節すれば過富最もtlましい結果が得ら
れる0かくして培養2日から10日後には薗体中訃よび
/又は培養液中に8ムMが生成蓄積するので、これを常
法に従って処理することによpaAMt*得することが
できるO例えば13AM含有曹体を過塩素置で抽出し、
抽出at氷冷下に炭酸水嵩力゛リクムで中和したのち、
必要に応じて強酸性カチオン交換@脂に接触させ、次い
で5Aiit−黴着しえのち硫酸で溶出し、溶出液にす
/タ/グステ/#1を加えて8ムMを沈澱させることに
よって単離することができる0以下に実施例を挙げて本
発明をさらに具体的に親羽するが、本発明はこれに限定
されるものではない・表お、実施例における8ムMの定
量は次のようにして行った・すなわち、培養終了後、遠
心分離にて菌体と培養液を分離し、菌体を約5倍量の1
.5M過塩素酸で抽出し九〇次いで得られた抽出上澄液
を培養液と混倉したのち、ベーノj−クロマトグラフィ
ー(展開溶媒:エタノール/1−ブタノール/水/酢酸
/1−ピロリン酸ナトリウム水II波−55/1G/I
O/1/1)で分離し、紫外−検出器で8AM相当のス
ポットを検出し、これを(11M塩酸で抽出して、その
2604懸の吸光度より試料中の8AMjlt−検出し
た。The yeast used in the present invention may be any yeast as long as it has the ability to accumulate flAM in a methionine-containing medium, but yeasts belonging to the genus Saccharomyces are preferred, and specific examples include Naccalomyces cerevisiae po 2546, Candida Φ ! Sedonye/Sis IFO0960, Ha/Zenuts Fabianii IP0 1
570 etc.'. tlI - Even these natural and artificial mutant bacteria can be used in the same manner as Kagishii, which has the ability to write BAM. It is essential to add lower peptides consisting of glycine and alanine. ・Lower peptides here refer to dipeptides, tripeptides, and tetrapeptides. Specific examples of such lower peptides include: For example, glycylglycine, glyculglycylglycine, glycylglycylglycine, D, L-Atzel D, L-Acunino, L-Atzunil L
-Afnyl L-72, L-alanyl L-alanyl L
- Alanyl L-792, L-72 Nylglycine, etc. - These O lower peptides are K(L
It is added at a concentration of O2#/jJ or more, especially cLo41
/dl or higher is appropriate.Other conditions for the culture medium used in the present invention may be determined according to conventional methods. Acids, hydrocarbons, alcohols such as methanol and ethanol, etc. can be used. Further, in the culture medium, ordinary nitrogen sources, inorganic salts, and organic micronutrients are used as necessary.For O 2 culture, aerobic conditions are preferable, and the culture temperature is preferably in the range of 20[11' to 40C]. During culture, adjusting the pH of the medium to a range of 3 to 8 will yield the most desirable results.Thus, after 2 to 10 days of culture, 8 mu. Since M is generated and accumulated, paAMt* can be obtained by processing it according to a conventional method.
After extraction at ice-cooling and neutralization with carbonated water bulk liquor,
If necessary, contact with strongly acidic cation exchange@fat, then 5Aiit-moulding, then elute with sulfuric acid, add Su/Ta/Guste/#1 to the eluate to precipitate 8mM. The present invention will be described in more detail with examples below, but the present invention is not limited thereto. After the cultivation was completed, the bacterial cells and the culture solution were separated by centrifugation, and the bacterial cells were diluted with approximately 5 times the volume of 1
.. After extraction with 5M perchloric acid, the resulting extraction supernatant was mixed with the culture solution, and then subjected to beno-J chromatography (developing solvent: ethanol/1-butanol/water/acetic acid/sodium 1-pyrophosphate). Water II wave-55/1G/I
A spot corresponding to 8AM was detected using an ultraviolet detector, and this was extracted with 11M hydrochloric acid, and 8AMjlt- in the sample was detected from the absorbance of 2604 times.
実施11i11
グルコース 5 lid J 、ポリペプトン [1L
5t/di、K H4F Os a41 /d J、
1cmMP Oa ’(L41 /d J、MgS、0
4−7H!0α021/d1%酵母エキス CL21/
d1. 寒天2#/dJからなる寒天斜面培地(PH
60)に2日間生育させたナツカロマイセス・セレビシ
ェIP’02546の1白金耳を、シュクロース 10
1/di。Run 11i11 Glucose 5 lid J, Polypeptone [1L
5t/di, K H4F Os a41 /d J,
1cmMP Oa'(L41/dJ, MgS, 0
4-7H! 0α021/d1% yeast extract CL21/
d1. Agar slant medium (PH
1 platinum loop of Natsucalomyces cerevisiae IP'02546 grown for 2 days on 60) was mixed with 10 sucrose.
1/di.
酵母エキス 1t7d s、 KH,PO2α41/d
i%Mg804・7H20α011/dj、尿素(別滅
劇) LSI/di。Yeast extract 1t7ds, KH, PO2α41/d
i%Mg804・7H20α011/dj, urea (separate play) LSI/di.
L−メチオニン Q、751/d j、 Oa c、t
2@ 2 a2゜α(J21/dj、Zo80a ・7
H20α25vml/dl、Fe3O4・7H20α2
5m1/lN、MnSO4・4〜6T120 t25
wsl/di、0u804 ” 5 H2O2μl/d
i、HmBOs 2μI/dl。L-methionine Q, 751/d j, Oa c, t
2 @ 2 a2゜α (J21/dj, Zo80a ・7
H20α25vml/dl, Fe3O4・7H20α2
5m1/lN, MnSO4・4~6T120 t25
wsl/di, 0u804” 5 H2O2μl/d
i, HmBOs 2μI/dl.
0oCI2 ・6 H2O0,2fil/di、 II
1fil/di及び所定量のグリシルグリシ/から
な5 PHlに調整、加熱滅菌した培地5dに植1し、
28Cで5日間振菫し九0法いで8ムMの蓄積量tm定
し、結果を#!1表に示し九〇
第 1 表
この結果から、グリシルグリシンを添加することKよ、
DaA)C)生成量を向上させうろことがわかる。0oCI2 ・6 H2O0,2fil/di, II
1 fil/di and a predetermined amount of glycylglycyl/karana 5 PHl adjusted to 5 d of heat-sterilized medium,
Shake at 28C for 5 days, determine the accumulated amount tm of 8M using the 90 method, and write the result as #! Table 1 shows Table 90. From this result, it is recommended to add glycylglycine.
It can be seen that the amount of DaA)C) produced can be improved.
実Jlall’12
グリシルグリシ/のかわりK111g2表に示す低級ペ
プチドをα11/Ill 09度て培地に添加すること
以外は実施例1と同じ条件で培養を行った。結果を第2
表に示した。Cultivation was carried out under the same conditions as in Example 1, except that the lower peptides shown in the K111g2 table were added to the medium at α11/Ill 09 degrees instead of K111g2. Second result
Shown in the table.
j[2表
*1 加熱Millせずに常温で無−的に培地に添加し
た。j[Table 2 *1 It was added to the culture medium without heating at room temperature without heating.
実施例5
ナツカロマイセス・セレビジエエP0 2346のかわ
)に菖3表に示す菌株を用いること及び第5表に示す低
級ペプチドを培地に添加すること以外は、実施例1と同
じ条件で培養を行なつ九。結果を第5表に示した〇
第5表Example 5 Cultivation was carried out under the same conditions as in Example 1, except that the strains shown in Table 3 were used for Natsucharomyces cerevisiae (P0 2346) and the lower peptides shown in Table 5 were added to the medium. . The results are shown in Table 5.〇Table 5
Claims (1)
チオニン含有培地で培養してS−アデノシルメチオニン
會贋造する方法において、グリシン及び/i丸はアラニ
ンから構成される低級ペプチドを培地に添加することt
Ili黴とする8−アデノシルメチオエ10Jll造方
法。 2、 l1m1カナツカロマイセス属、キャ/デイダ
属を九はハ/ゼXう属に属するものである特許請求aS
S鎮1項記載の方法。[Claims] tS-adenosylmethioni/Ikudo #! In the method for producing S-adenosylmethionine by culturing yeast having methionine in a methionine-containing medium, a lower peptide consisting of glycine and alanine is added to the medium.
A method for producing 10 Jll of 8-adenosylmethioe as Ili mold. 2. A patent claim aS that belongs to the genus Ha/Deida and the genus I1m1
The method described in Section 1 of S.
Priority Applications (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP13456181A JPS5836397A (en) | 1981-08-27 | 1981-08-27 | Preparation of s-adenosyl methionine |
CH5083/82A CH648864A5 (en) | 1981-08-27 | 1982-08-26 | Process for the preparation of S-adenosylmethionine |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP13456181A JPS5836397A (en) | 1981-08-27 | 1981-08-27 | Preparation of s-adenosyl methionine |
Publications (2)
Publication Number | Publication Date |
---|---|
JPS5836397A true JPS5836397A (en) | 1983-03-03 |
JPH0153037B2 JPH0153037B2 (en) | 1989-11-10 |
Family
ID=15131205
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP13456181A Granted JPS5836397A (en) | 1981-08-27 | 1981-08-27 | Preparation of s-adenosyl methionine |
Country Status (1)
Country | Link |
---|---|
JP (1) | JPS5836397A (en) |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6881837B2 (en) | 2001-06-07 | 2005-04-19 | Orchid Chemicals & Pharmaceuticals Ltd. | Chemical synthesis of S-adenosyl-L-methionine with enrichment of (S,S)-isomer |
US7667034B2 (en) | 2004-06-23 | 2010-02-23 | Orchid Chemicals & Pharmaceuticals Limited | Chemical synthesis of S-adenosyl-L-methionine with enrichment of (S,S)-isomer |
JP2011177125A (en) * | 2010-03-02 | 2011-09-15 | National Research Inst Of Brewing | Method for obtaining highly s-adenosylmethionine-accumulating yeast |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS5082288A (en) * | 1973-11-27 | 1975-07-03 |
-
1981
- 1981-08-27 JP JP13456181A patent/JPS5836397A/en active Granted
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JPS5082288A (en) * | 1973-11-27 | 1975-07-03 |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6881837B2 (en) | 2001-06-07 | 2005-04-19 | Orchid Chemicals & Pharmaceuticals Ltd. | Chemical synthesis of S-adenosyl-L-methionine with enrichment of (S,S)-isomer |
US7667034B2 (en) | 2004-06-23 | 2010-02-23 | Orchid Chemicals & Pharmaceuticals Limited | Chemical synthesis of S-adenosyl-L-methionine with enrichment of (S,S)-isomer |
JP2011177125A (en) * | 2010-03-02 | 2011-09-15 | National Research Inst Of Brewing | Method for obtaining highly s-adenosylmethionine-accumulating yeast |
Also Published As
Publication number | Publication date |
---|---|
JPH0153037B2 (en) | 1989-11-10 |
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