JPS5841898A - Production of s-adenosyl-l-homocysteine - Google Patents
Production of s-adenosyl-l-homocysteineInfo
- Publication number
- JPS5841898A JPS5841898A JP14146381A JP14146381A JPS5841898A JP S5841898 A JPS5841898 A JP S5841898A JP 14146381 A JP14146381 A JP 14146381A JP 14146381 A JP14146381 A JP 14146381A JP S5841898 A JPS5841898 A JP S5841898A
- Authority
- JP
- Japan
- Prior art keywords
- sam
- adenosyl
- reaction
- sah
- thiosulfate
- 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.)
- Pending
Links
Landscapes
- Saccharide Compounds (AREA)
Abstract
Description
【発明の詳細な説明】
本発明はS−アデノシル−L−メチオニン(以下SAM
と略称する。)の化学的な脱メチル化によるS−アデノ
シル−L−ホモシスティン(以下SAHと略称する。)
の製造法に関する。DETAILED DESCRIPTION OF THE INVENTION The present invention relates to S-adenosyl-L-methionine (hereinafter SAM
It is abbreviated as. ) by chemical demethylation of S-adenosyl-L-homocysteine (hereinafter abbreviated as SAH).
Concerning the manufacturing method.
SAHは、生体内で普遍的なSAMを基質とするトラン
スメチル化反応で生成する重要な生物学的中間物質であ
り、S A、MのトランスメチJし化反応の生成物阻害
物質として知られている。近年、SAHそれ自体の薬理
活性が注目され、中枢神経系統の鎮静作用および睡眠誘
発作用、ならびに抗痙彎作用等が報告されている(特開
昭54−145222号公報参照)。SAH is an important biological intermediate produced in the transmethylation reaction using SAM as a substrate, which is common in living organisms, and is known as a product inhibitor of the transmethylation reaction of SA and M. There is. In recent years, the pharmacological activity of SAH itself has attracted attention, and its sedative and sleep-inducing effects on the central nervous system, as well as its anti-spasmodic effects, etc. have been reported (see JP-A-54-145222).
ジ
SAHの製造法としては、tlls−ベンゼルーL−ホ
モシスティンとトシルイソプロピナデンアデノシンを原
料とする化学合成法(Biochem。As a method for producing di-SAH, a chemical synthesis method (Biochem) using tlls-benzel-L-homocystine and tosylisopropinadenadenosine as raw materials is used.
Prel)arations、 VOl、 8 、1)
p、 8−11 (1961)参照)、(2)酵母菌体
から過塩素酸によってSAMとともに抽出し、SAHを
分離する方法(Anal、 Biochem、。Prel)arations, VOl, 8, 1)
p. 8-11 (1961)), (2) A method of extracting SAH from yeast cells with perchloric acid together with SAM (Anal, Biochem, 2003).
Vol、 15 、 I)I)、 828−888 (
19661参照) 、+81アデノシンとホモシスティ
ンを原料として、ラット等の肝臓から抽出した酵素によ
って酵素合成する方法(J、Biol、(hem、、V
ol、 284 、1)l)、 60 B−60,8(
1959)参照) 、 (41酵母菌体から抽出、精製
したメチルトランスフェラーゼを用い、ホモシスティン
とSAMを基質として反応させ、メチオニンとともにS
AHを生成させる方法(J。Vol, 15, I)I), 828-888 (
19661), a method of enzymatic synthesis using enzymes extracted from the liver of rats etc. using +81 adenosine and homocysteine as raw materials (J, Biol, (hem, , V
ol, 284, 1)l), 60 B-60,8(
(1959)), (41) Using methyltransferase extracted and purified from yeast cells, homocysteine and SAM were reacted as substrates, and methionine and S
Method for producing AH (J.
Biol、(:hem、、Vol、289.pI)、
1551−1556 (1964)参照)、(51sA
Mのギ酸塩、酢酸塩、過塩素酸塩、塩酸塩、臭化水素酸
塩、沃化水素酸剤
塩などの使用有機溶損に溶解する特定の塩を、特定の一
性有機溶剤またはその混合溶剤に溶解し、脱水した後、
沃化バリウム、沃化ナトリウムなどのアルカリ金属もし
くはアルカリ土類金属のハロゲン化物またはブソイドハ
ロゲニドと反応させ、SAMを脱メチル化してSAHを
製造する方法(5
特公昭4F=* −87586号公報参照)などが知ら
れている。Biol, (:hem,, Vol, 289.pI),
1551-1556 (1964)), (51sA
Formate, acetate, perchlorate, hydrochloride, hydrobromide, hydroiodide salt, etc. of M After dissolving in mixed solvent and dehydrating,
A method for producing SAH by demethylating SAM by reacting it with an alkali metal or alkaline earth metal halide such as barium iodide or sodium iodide or a butoid halide (5 Japanese Patent Publication No. 4F=*-87586) ) are known.
しかしながら、(1)の方法は、液体アンモニアを使用
するため、反応系を高圧、低温に保たなければならず、
また、乾燥窒素ガス中で反応するなど特殊な設備および
条件を必要とする。さらに、原料化合物の保護および生
成物の脱保護などの工程が必要であり、工業的生産には
適さない方法である。(2)の方法は、酵母菌体中のS
AHが微量て、効率が悪く、また、抽出液からのSAH
の分離が困難である。]3)の方法は、動物の肝臓から
抽出した酵素を使用するため、工業的生産には採用てき
ない。(4)の方法は、酵母菌体から酵素を抽出する8
工程が煩雑であり、収率も悪い。(5)の方法は、上記
の方法に比べると現実的な方法であるが、反応を゛無水
の極性有機溶媒中で行う必要があり、本来。However, since method (1) uses liquid ammonia, the reaction system must be kept at high pressure and low temperature.
In addition, special equipment and conditions are required, such as reaction in dry nitrogen gas. Furthermore, steps such as protection of the raw material compound and deprotection of the product are required, making the method unsuitable for industrial production. Method (2) is based on S in yeast cells.
There is a small amount of AH, the efficiency is poor, and SAH from the extract
is difficult to separate. The method 3) cannot be used for industrial production because it uses enzymes extracted from animal livers. Method (4) involves extracting enzymes from yeast cells.
The process is complicated and the yield is poor. Method (5) is a more realistic method than the above method, but the reaction must be carried out in an anhydrous polar organic solvent.
水との親和性が強いSAMの溶液を脱水するための複雑
な工程が必要となる。また、このため原料となるSAM
の塩の形態も限られる。A complicated process is required to dehydrate a solution of SAM, which has a strong affinity for water. In addition, for this reason, the raw material SAM
The form of the salt is also limited.
本発明者らは、これら従来技術の欠点を克服し、容易に
入手できるSAMを原料とし、取扱いの容易な反応試薬
を使用し、水溶液中で簡単な設備および比較的温和な条
件で反応することにより、従段
未決に比べ、格榊の収率てSAHを製造することに成功
し、本発明を完成した。The present inventors have overcome the drawbacks of these conventional techniques, used easily available SAM as a raw material, used easy-to-handle reaction reagents, and carried out the reaction in an aqueous solution with simple equipment and relatively mild conditions. As a result, we succeeded in producing SAH with a higher yield than the previous method, and completed the present invention.
本発明は、SAMの脱メチル化によってSAWを製造す
るに際し、SAMの水溶性塩を゛水溶液中でチオ硫酸塩
、亜硫酸塩、亜硫酸水素塩またはピロ亜硫酸塩と反応さ
せて脱メチル化することを特徴とするSAHの製造法を
提供するものである、本発明において原料として使用さ
れるSAMは水溶性の塩であればよく、特定の塩に限定
されない。たとえば、塩酸塩、硫酸塩、沃化水素酸塩、
臭化水素酸塩、りん酸塩、過塩素酸塩などの無機酸塩、
またはギ酸塩、酢酸塩、クエン酸塩、≠7酒石酸塩、リ
ンゴ酸塩、コハク酸塩、有機スルホン酸塩(たとえば、
トルエンスルホン酸塩など)、ヌクレオシド硫酸エステ
ル塩、糖硫酸エステル塩などの有機酸塩などいずれも使
用できる。またSAMの塩は粗精製のものであってもよ
く、たとえば酵母抽出液からの部分精製物であってもよ
い。In producing SAW by demethylation of SAM, the present invention involves demethylating a water-soluble salt of SAM with thiosulfate, sulfite, bisulfite or pyrosulfite in an aqueous solution. SAM used as a raw material in the present invention, which provides a characteristic method for producing SAH, may be any water-soluble salt and is not limited to a specific salt. For example, hydrochloride, sulfate, hydroiodide,
inorganic acid salts such as hydrobromide, phosphate, perchlorate;
or formates, acetates, citrates, ≠7tartrates, malates, succinates, organic sulfonates (e.g.
Toluene sulfonate, etc.), organic acid salts such as nucleoside sulfate ester salts, sugar sulfate ester salts, etc. can be used. Further, the SAM salt may be crudely purified, for example, a partially purified product from yeast extract.
脱メチル化反応の反応試薬としてはチオ硫酸ナトリウム
、チオ硫酸アンモニウム、チオ硫酸カリウム、チオ硫酸
カルシウム、チオ硫酸マグネシウムなどのチオ硫酸塩、
亜硫酸ナトリウム、亜硫酸カリウム、亜硫酸リチウム、
亜硫酸アンモニウムなどの亜硫酸塩、亜硫酸水素ナトリ
ウム、亜硫酸水素カリウムなどの亜硫酸水素塩、または
ピロ亜硫酸ナトリウム、ピロ亜硫酸カリウムなどのピロ
亜硫酸塩が挙げられる。As reaction reagents for the demethylation reaction, thiosulfates such as sodium thiosulfate, ammonium thiosulfate, potassium thiosulfate, calcium thiosulfate, and magnesium thiosulfate;
Sodium sulfite, potassium sulfite, lithium sulfite,
Examples include sulfites such as ammonium sulfite, hydrogen sulfites such as sodium hydrogen sulfite and potassium hydrogen sulfite, and pyrosulfites such as sodium pyrosulfite and potassium pyrosulfite.
本発明の反応は水溶液中で行なわれる。反応を水溶液中
で行うことにより、従来技術Jζ比へて原料選択の範囲
が広がり、反応操作および精製操作が簡単となるなど種
々の利点が生じた。反応条件は特+(限定されないが、
実用的条件はpH1,0〜7.o。The reaction of the present invention is carried out in an aqueous solution. By carrying out the reaction in an aqueous solution, various advantages such as a wider range of raw material selection than the Jζ ratio of the prior art and simplification of reaction and purification operations were produced. The reaction conditions are specified (but not limited to)
Practical conditions are pH 1.0-7. o.
温度は室温〜60℃である。すなわち、pHが低すぎる
場合または温度が低すぎる場合には反応は好適に進行せ
ず、pHが高すぎる場合または温度が高すぎる場合には
SAMの分解、物であるメチルチオアデノシンおよびア
デニンが増加し、SAHの収率が低下する。好適な反応
条件はpH8〜6、温度30〜50℃である。反応試薬
の量も特に限定されないがSAMの塩に対して10倍当
量以下、好ましくは2〜5倍当量を使用すれば経済的で
ある。The temperature is room temperature to 60°C. That is, if the pH is too low or the temperature is too low, the reaction will not proceed properly, and if the pH is too high or the temperature is too high, SAM will decompose and the products methylthioadenosine and adenine will increase. , the yield of SAH decreases. Suitable reaction conditions are pH 8-6 and temperature 30-50°C. The amount of the reaction reagent is also not particularly limited, but it is economical to use an amount of 10 times the equivalent or less, preferably 2 to 5 times the equivalent of the SAM salt.
反応液からのSAHの分離精製は常法により好適に行な
われる。すなわち、活性炭吸着法、弱酸性陽イオン交換
樹脂、強酸性陽イオン交換樹脂などによるイオン交換ク
ロマトグラフ法、有機溶媒、温度差などを利用した結晶
化法など、その他SAHの精製に通常利用される方法は
いずれも本発明における反応液からのSAHの分離精製
に適宜採用される。Separation and purification of SAH from the reaction solution is suitably carried out by conventional methods. That is, activated carbon adsorption method, ion exchange chromatography method using weakly acidic cation exchange resin, strong acid cation exchange resin, etc., crystallization method using organic solvent, temperature difference, etc., and other methods commonly used to purify SAH. Any of these methods can be appropriately employed for separating and purifying SAH from the reaction solution in the present invention.
以下、本発明を実施例によって具体的に説明するが、本
発明はこれに限定されるものではない。EXAMPLES Hereinafter, the present invention will be specifically explained with reference to Examples, but the present invention is not limited thereto.
実施例1
粗精製されたSAM硫酸塩50 g(SAM純度58.
4%)を水400 mlに溶解し、チオ硫酸ナトリウム
589を添加した後、水酸化ナトリウムで1)Hを4.
5に調整した。この水溶液を50°Cで撹拌しながら7
時間反応を行った。Example 1 50 g of crudely purified SAM sulfate (SAM purity 58.
4%) in 400 ml of water, and after adding 589 ml of sodium thiosulfate, 1)H was dissolved in 4% with sodium hydroxide.
Adjusted to 5. While stirring this aqueous solution at 50°C,
A time reaction was performed.
反応液を濾過後、1000 mlに希釈してダイヤイオ
ンWK−20(商品名、三菱化成工業■製)H型500
mlに1000g//時間で吸着した。水洗後、00
1N塩酸で溶出し、IN水酸化ナトリウムで中和後、減
圧下で150 mlに濃縮し、エタノール10100O
を加えて一夜冷却放置し、SAHの結晶を得た。この結
晶を少量の氷水で洗浄後、熱水より再結晶させ、SAH
・2 H2Oの結晶を24.9g得た(SAMよりの理
論モル収率81.196)。After filtering the reaction solution, it was diluted to 1000 ml and diluted with Diaion WK-20 (trade name, manufactured by Mitsubishi Chemical Corporation) H type 500.
ml at a rate of 1000 g/hour. After washing with water, 00
Elute with 1N hydrochloric acid, neutralize with IN sodium hydroxide, concentrate to 150 ml under reduced pressure, and add ethanol 10100O
was added and left to cool overnight to obtain crystals of SAH. After washing the crystals with a small amount of ice water, they were recrystallized from hot water and SAH
24.9 g of 2 H2O crystals were obtained (theoretical molar yield from SAM: 81.196).
実施例2
ジュレンクツ培地(J、 Biol、 (hem、 、
Vol、 229 。Example 2 Jurenkz medium (J, Biol, (hem, ,
Vol, 229.
p、 1087 (1957)参照)で30°Cl2O
時間培養した市販パン酵母(オリエンタル酵母工業(抹
製)1.0#よりIN塩酸で抽出して中和後、H型に調
整したダイヤイオンWK−,10(商品名。p, 1087 (1957)) at 30°Cl2O
Diaion WK-, 10 (trade name) was extracted from commercially available baker's yeast (Oriental Yeast Kogyo Co., Ltd. (Maga) 1.0#) cultured for a period of time with IN hydrochloric acid, neutralized, and adjusted to H type.
三菱化成工業(株制)1011こ吸着、水洗後、01N
塩酸で溶出し、9.49のSAMを塩酸塩として含んだ
溶液20eを得た。この溶液をpH5,0に調整し、チ
オ硫酸ナトリウム200q添加後、400Cて10時間
撹拌放置した。Mitsubishi Chemical Industries, Ltd. 1011 adsorption, after washing with water, 01N
Elution was carried out with hydrochloric acid to obtain a solution 20e containing 9.49 SAM as a hydrochloride. This solution was adjusted to pH 5.0, and after adding 200q of sodium thiosulfate, it was left stirring at 400C for 10 hours.
この溶液を800 mlの活性炭カラムに通し、蒸留水
で水洗後、エタノール−水−濃アンモニア(50:50
:1)で、溶出し、257nmの紫外線吸収画分80
00 mlを得た。この溶液を20 mlに濃縮後、中
和して一夜08Cで放置してSAHを析出させた。結晶
を濾過分離し、少量の氷水で洗浄後、熱水より再結晶さ
せてSAH・2 H2Oの結晶6.59を得た(SAM
よりの理論モル収率67.3%)。This solution was passed through an 800 ml activated carbon column, washed with distilled water, and then mixed with ethanol-water-concentrated ammonia (50:50).
:1), eluted, 257 nm ultraviolet absorption fraction 80
00 ml was obtained. After concentrating this solution to 20 ml, it was neutralized and left overnight at 08C to precipitate SAH. The crystals were separated by filtration, washed with a small amount of ice water, and then recrystallized from hot water to obtain 6.59 crystals of SAH.2H2O (SAM
(theoretical molar yield of 67.3%).
実施例8
粗精製されたSAM硫酸塩10g(SAM純度5864
%)を100 mlの水に溶解後、チオ硫酸アンモニウ
ム122gを添加し、水酸化ナトリウムでpHを451
こ調整後、50°Cで7時間撹拌反応させた後、実施例
1と同様番こ精製してSAH・2H20の結晶4.8g
を得た(SAMよりの理論収率78%)。Example 8 10 g of crudely purified SAM sulfate (SAM purity 5864
%) in 100 ml of water, 122 g of ammonium thiosulfate was added, and the pH was adjusted to 451 with sodium hydroxide.
After this adjustment, the reaction was stirred at 50°C for 7 hours, and then purified in the same manner as in Example 1 to obtain 4.8 g of crystals of SAH・2H20.
(78% theoretical yield from SAM).
実施例4
粗精製されたSAM硫酸塩10g(SAM純度584%
)を100 mlの水に溶解した後、亜硫酸ナトリウム
5.58gを添加し、水酸化ナトリウムでpHを5.0
に調整後、50’Cて6時間撹拌した後、実施例1と同
様に精製してSAH・2H20の結晶を2.1g得た(
SAMよりの理論モル収率84.0%)。Example 4 10 g of crudely purified SAM sulfate (SAM purity 584%)
) was dissolved in 100 ml of water, 5.58 g of sodium sulfite was added, and the pH was adjusted to 5.0 with sodium hydroxide.
After adjusting to
(Theoretical molar yield from SAM 84.0%).
特許出願人 (677)ヤマサ醤油株式会社101Patent applicant (677) Yamasa Soy Sauce Co., Ltd. 101
Claims (1)
S−アデノシル−L−ホモシスティンを製造するに際し
、S−アデノシル−し−メチオニンの水溶性塩を水溶液
中でチオ硫酸塩、亜硫酸塩、亜硫酸水素塩またはピロ亜
硫酸塩と反応させて脱メチル化することを特徴とするS
−アゾノンルーム−ホモシスティンの製造法。When producing S-adenosyl-L-homocysteine by demethylation of S-adenosyl-methionine, the water-soluble salt of S-adenosyl-methionine is converted into thiosulfate, sulfite, and bisulfite in an aqueous solution. or S characterized by demethylation by reaction with pyrosulfite
- Azononroom - Method for producing homocystine.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP14146381A JPS5841898A (en) | 1981-09-07 | 1981-09-07 | Production of s-adenosyl-l-homocysteine |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP14146381A JPS5841898A (en) | 1981-09-07 | 1981-09-07 | Production of s-adenosyl-l-homocysteine |
Publications (1)
Publication Number | Publication Date |
---|---|
JPS5841898A true JPS5841898A (en) | 1983-03-11 |
Family
ID=15292464
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP14146381A Pending JPS5841898A (en) | 1981-09-07 | 1981-09-07 | Production of s-adenosyl-l-homocysteine |
Country Status (1)
Country | Link |
---|---|
JP (1) | JPS5841898A (en) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5114931A (en) * | 1983-08-24 | 1992-05-19 | Bioresearch S.P.A. | Injectable therapeutic compositions containing stable s-adenosyl-l-methionine salts |
-
1981
- 1981-09-07 JP JP14146381A patent/JPS5841898A/en active Pending
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5114931A (en) * | 1983-08-24 | 1992-05-19 | Bioresearch S.P.A. | Injectable therapeutic compositions containing stable s-adenosyl-l-methionine salts |
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