JPH0338840B2 - - Google Patents
Info
- Publication number
- JPH0338840B2 JPH0338840B2 JP18653084A JP18653084A JPH0338840B2 JP H0338840 B2 JPH0338840 B2 JP H0338840B2 JP 18653084 A JP18653084 A JP 18653084A JP 18653084 A JP18653084 A JP 18653084A JP H0338840 B2 JPH0338840 B2 JP H0338840B2
- Authority
- JP
- Japan
- Prior art keywords
- valine
- acetyl
- isopropylhydantoin
- isolated
- reaction solution
- 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.)
- Expired
Links
- KZSNJWFQEVHDMF-BYPYZUCNSA-N L-valine Chemical compound CC(C)[C@H](N)C(O)=O KZSNJWFQEVHDMF-BYPYZUCNSA-N 0.000 claims description 100
- 229960004295 valine Drugs 0.000 claims description 58
- KZSNJWFQEVHDMF-UHFFFAOYSA-N Valine Chemical compound CC(C)C(N)C(O)=O KZSNJWFQEVHDMF-UHFFFAOYSA-N 0.000 claims description 51
- IHYJTAOFMMMOPX-ZCFIWIBFSA-N (2r)-2-acetamido-3-methylbutanoic acid Chemical compound CC(C)[C@H](C(O)=O)NC(C)=O IHYJTAOFMMMOPX-ZCFIWIBFSA-N 0.000 claims description 21
- KZSNJWFQEVHDMF-SCSAIBSYSA-N D-valine Chemical compound CC(C)[C@@H](N)C(O)=O KZSNJWFQEVHDMF-SCSAIBSYSA-N 0.000 claims description 21
- 229930182831 D-valine Natural products 0.000 claims description 21
- 238000000034 method Methods 0.000 claims description 19
- LTJKEWJBNJGKCM-UHFFFAOYSA-N 1-propan-2-ylimidazolidine-2,4-dione Chemical compound CC(C)N1CC(=O)NC1=O LTJKEWJBNJGKCM-UHFFFAOYSA-N 0.000 claims description 18
- 239000000243 solution Substances 0.000 claims description 18
- 238000006243 chemical reaction Methods 0.000 claims description 17
- 239000004474 valine Substances 0.000 claims description 15
- IHYJTAOFMMMOPX-UHFFFAOYSA-N N-Acetyl-Valine Chemical compound CC(C)C(C(O)=O)NC(C)=O IHYJTAOFMMMOPX-UHFFFAOYSA-N 0.000 claims description 12
- 230000002378 acidificating effect Effects 0.000 claims description 11
- NWUYHJFMYQTDRP-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenyl-2-ethylbenzene;styrene Chemical compound C=CC1=CC=CC=C1.CCC1=CC=CC=C1C=C.C=CC1=CC=CC=C1C=C NWUYHJFMYQTDRP-UHFFFAOYSA-N 0.000 claims description 10
- 239000000706 filtrate Substances 0.000 claims description 10
- 238000005904 alkaline hydrolysis reaction Methods 0.000 claims description 6
- 238000004519 manufacturing process Methods 0.000 claims description 6
- 239000003729 cation exchange resin Substances 0.000 claims description 5
- 230000003301 hydrolyzing effect Effects 0.000 claims description 5
- 238000006640 acetylation reaction Methods 0.000 claims description 3
- 239000007864 aqueous solution Substances 0.000 claims description 3
- 238000002955 isolation Methods 0.000 claims description 3
- 239000007790 solid phase Substances 0.000 claims description 3
- 125000000738 acetamido group Chemical group [H]C([H])([H])C(=O)N([H])[*] 0.000 claims description 2
- 230000021736 acetylation Effects 0.000 claims description 2
- 238000003916 acid precipitation Methods 0.000 claims description 2
- 238000010828 elution Methods 0.000 claims description 2
- 238000005903 acid hydrolysis reaction Methods 0.000 claims 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 24
- WFDIJRYMOXRFFG-UHFFFAOYSA-N Acetic anhydride Chemical compound CC(=O)OC(C)=O WFDIJRYMOXRFFG-UHFFFAOYSA-N 0.000 description 18
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 15
- IHYJTAOFMMMOPX-LURJTMIESA-N N-acetyl-L-valine Chemical compound CC(C)[C@@H](C(O)=O)NC(C)=O IHYJTAOFMMMOPX-LURJTMIESA-N 0.000 description 10
- 239000013078 crystal Substances 0.000 description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 9
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 8
- 239000000203 mixture Substances 0.000 description 7
- 230000003287 optical effect Effects 0.000 description 7
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 6
- 238000006460 hydrolysis reaction Methods 0.000 description 6
- 239000007788 liquid Substances 0.000 description 6
- 238000001914 filtration Methods 0.000 description 5
- 239000003456 ion exchange resin Substances 0.000 description 5
- 229920003303 ion-exchange polymer Polymers 0.000 description 5
- 238000005406 washing Methods 0.000 description 5
- 239000003513 alkali Substances 0.000 description 4
- 229940024606 amino acid Drugs 0.000 description 4
- 150000001413 amino acids Chemical class 0.000 description 4
- 239000011780 sodium chloride Substances 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 238000001035 drying Methods 0.000 description 3
- 238000000855 fermentation Methods 0.000 description 3
- 230000004151 fermentation Effects 0.000 description 3
- 230000007062 hydrolysis Effects 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 238000010992 reflux Methods 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 229910001415 sodium ion Inorganic materials 0.000 description 3
- 238000003756 stirring Methods 0.000 description 3
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 2
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 235000011114 ammonium hydroxide Nutrition 0.000 description 2
- 238000007796 conventional method Methods 0.000 description 2
- 238000001816 cooling Methods 0.000 description 2
- 238000002425 crystallisation Methods 0.000 description 2
- 230000008025 crystallization Effects 0.000 description 2
- 239000003480 eluent Substances 0.000 description 2
- BDERNNFJNOPAEC-UHFFFAOYSA-N propan-1-ol Chemical compound CCCO BDERNNFJNOPAEC-UHFFFAOYSA-N 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- UQDJGEHQDNVPGU-UHFFFAOYSA-N serine phosphoethanolamine Chemical compound [NH3+]CCOP([O-])(=O)OCC([NH3+])C([O-])=O UQDJGEHQDNVPGU-UHFFFAOYSA-N 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 229960000583 acetic acid Drugs 0.000 description 1
- 230000000397 acetylating effect Effects 0.000 description 1
- 239000003905 agrochemical Substances 0.000 description 1
- 150000001298 alcohols Chemical class 0.000 description 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-N ammonia Natural products N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 108091022884 dihydropyrimidinase Proteins 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 235000020776 essential amino acid Nutrition 0.000 description 1
- 239000003797 essential amino acid Substances 0.000 description 1
- 239000012362 glacial acetic acid Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
Description
【発明の詳細な説明】
産業上の利用分野
本発明は、イソプロピルヒダントインからＤ−
バリン、及びＬ−バリンを同時に製造する方法に
関する。DETAILED DESCRIPTION OF THE INVENTION Field of Industrial Application The present invention is directed to the production of isopropyl hydantoin from D-
The present invention relates to a method for simultaneously producing valine and L-valine.
さらに詳しくは、イソプロピルヒダントインを
アルカリ加水分解して得られたDL−バリンを含
む反応液に無水酢酸を作用させて、アセチル化反
応を行つたのち酸析してＮ−アセチル−DL−バ
リンの結晶を得、これをアシラーゼを用いてＮ−
アセチル−Ｌ−バリンのみを選択的に加水分解し
て得られた、主にＮ−アセチル−Ｄ−バリンとＬ
−バリンからなるアシラーゼ反応液からＤ−バリ
ン及びＬ−バリンを単離する方法に関する。 More specifically, acetic anhydride is applied to a reaction solution containing DL-valine obtained by alkaline hydrolysis of isopropylhydantoin to perform an acetylation reaction, followed by acid precipitation to crystallize N-acetyl-DL-valine. This was converted into N- using acylase.
Mainly N-acetyl-D-valine and L-valine obtained by selectively hydrolyzing only acetyl-L-valine.
- A method for isolating D-valine and L-valine from an acylase reaction solution containing valine.
Ｄ−バリンはアミノ酸の一つであり、抗生物質
などの医薬品原料、農薬中間体として有用な化合
物であり、またＬ−バリンは必須アミノ酸の一つ
であり両者ともに大きな商品価値を有する。 D-valine is one of the amino acids, and is a compound useful as a raw material for pharmaceuticals such as antibiotics and as an intermediate for agricultural chemicals, and L-valine is one of the essential amino acids, and both have great commercial value.
従来の技術
通常、Ｌ−バリンの製造方法は発酵方法による
製造方法が多数知られており（特公昭56−19234
外）、またＤ−バリンに関してもイソプロピルヒ
ダントインにある種の微生物が生産する酵素ヒダ
ントイナーゼを作用させて酵素的にＤ−バリンを
得る方法（発酵と工業Vol38 No.10p937外）など
若干知られている。PRIOR TECHNOLOGY There are many known methods for producing L-valine using fermentation methods (Special Publication No. 56-19234
Regarding D-valine, there are some known methods, such as a method for enzymatically obtaining D-valine by allowing isopropylhydantoin to act on the enzyme hydantoinase produced by a certain type of microorganism (Fermentation and Industry Vol. 38, No. 10, p. 937). .
これら発酵法や酵素法による製造方法では、い
ずれも光学活性体の一方のみしか得られず、同一
原料からともに有用なＤ体、Ｌ体の光学活性なバ
リンを得ることはできない。 These fermentation methods and enzymatic production methods yield only one of the optically active forms, and it is not possible to obtain both the useful D-form and L-form optically active valine from the same raw material.
これに対し、化学的方法による場合、DL−バ
リンがラセミ体として得られるので、これより通
常のアミノ酸の光学分割の常法にしたがい両者を
効率よく分離せねばならない。 On the other hand, when using a chemical method, DL-valine is obtained as a racemate, and the two must be efficiently separated from each other according to the conventional optical resolution method for amino acids.
化学的方法によるDL−バリンの製造方法の一
つとして、イソプロピルヒダントインをアルカリ
の存在下加水分解してDL−バリンのラセミ体を
得る方法（USP2480644）は公知であり、また
DL−バリンを単離後、氷酢酸を溶媒にして無水
酢酸でアセチル化してＮ−アセチル−DL−バリ
ンを得る方法（Chemistry of amino acid 1961
年刊、p2375）が知られている。 As one method for producing DL-valine by a chemical method, a method for obtaining a racemic form of DL-valine by hydrolyzing isopropylhydantoin in the presence of an alkali (USP 2480644) is known.
After isolating DL-valine, it is acetylated with acetic anhydride using glacial acetic acid as a solvent to obtain N-acetyl-DL-valine (Chemistry of amino acid 1961
Annual publication, p2375) is known.
発明が解決しようとする問題点
イソプロピルヒダントインの加水分解反応によ
り得られたラセミ体のDL−バリンを含む反応液
からＤ−バリン及びＬ−バリンを夫々分離する方
法として、通常アミノ酸の光学分割法の一つとし
て知られているように、ラセミ体を含む反応液を
アセチル化し、引続きＬ−バリンアセチル化物の
み加水分解するアシラーゼを作用させ、分割反応
終了後のＬ−バリンの析出した反応マスからＬ−
バリンを先に単離して、一方Ｎ−アセチル−Ｄ−
バリンを主に含む濾液から、Ｎ−アセチル−Ｄ−
バリンを晶出単離させ、これを加水分解してＤ−
バリンを得る方法も考えられる。Problems to be Solved by the Invention As a method for separating D-valine and L-valine from a reaction solution containing racemic DL-valine obtained by the hydrolysis reaction of isopropylhydantoin, optical resolution of amino acids is usually used. As is known, a reaction solution containing a racemate is acetylated, followed by the action of acylase that hydrolyzes only the acetylated L-valine, and L-valine is extracted from the reaction mass in which L-valine is precipitated after the completion of the split reaction. −
Valine was isolated first while N-acetyl-D-
From the filtrate mainly containing valine, N-acetyl-D-
Valine was crystallized and isolated, and this was hydrolyzed to give D-
Another possible method is to obtain valine.
しかしながら、この場合は、濾液中に若干の未
反応のＮ−アセチル−Ｌ−バリンと溶解度の大き
いＬ−バリンがＮ−アセチル−Ｄ−バリンに対し
通常30％以上も含有されているため、Ｄ−バリン
の精製工程において含有されるＬ−バリンを充分
淘汰できず高純度Ｄ−バリンを得ることがきわめ
て困難である上、収率も大幅に低下するし、同時
にＬ−バリンの損失をも伴う。 However, in this case, the filtrate contains some unreacted N-acetyl-L-valine and highly soluble L-valine, which usually accounts for 30% or more of N-acetyl-D-valine. - In the valine purification process, L-valine contained in it cannot be sufficiently removed, making it extremely difficult to obtain high-purity D-valine, and the yield is also significantly reduced, accompanied by loss of L-valine. .
問題を解決するための手段
本発明者らは、イソプロピルヒダントインから
DL−バリンを経てＤ−バリン及びＬ−バリンを
同時に、かつ工業的有利に製造する方法について
鋭意検討した結果、イソプロピルヒダントインの
アルカリ加水分解反応液中のDL−バリンをアセ
チル化したのちアシラーゼを作用させることによ
つて得られるＮ−アセチル−Ｄ−バリンとＬ−バ
リンを含有する反応液のPHを１以下に調整すれ
ば、両者の溶解度の相違により、Ｎ−アセチル−
Ｄ−バリンのみが固相分として析出し、Ｌ−バリ
ンは完全に溶解することを見出した。したがつ
て、析出したＮ−アセチル−Ｄ−バリンを濾別
し、酸性下で加水分解後、簡単な精製工程に付す
ことにより高純度のＤ−バリンを収率よく得るこ
とができる。Means for solving the problem The inventors have discovered that from isopropylhydantoin
As a result of intensive studies on a method for simultaneously and industrially advantageous production of D-valine and L-valine via DL-valine, we found that DL-valine in the alkaline hydrolysis reaction solution of isopropylhydantoin was acetylated and then acylase was applied. If the pH of the reaction solution containing N-acetyl-D-valine and L-valine obtained by
It was found that only D-valine was precipitated as a solid phase component, and L-valine was completely dissolved. Therefore, by filtering the precipitated N-acetyl-D-valine, hydrolyzing it under acidic conditions, and subjecting it to a simple purification step, highly pure D-valine can be obtained in good yield.
一方、Ｎ−アセチル−Ｄ−バリンを分離する際
に得られる、主成分としてＬ−バリンを含有する
濾液は強酸性陽イオン交換樹脂と接触させてＬ−
バリンを吸着させたのち、常法により溶離させる
ことにより、高純度のＬ−バリンが得られること
もわかり、本発明は完成されたものである。 On the other hand, the filtrate containing L-valine as a main component, which is obtained when separating N-acetyl-D-valine, is brought into contact with a strongly acidic cation exchange resin.
It was also found that highly pure L-valine can be obtained by adsorbing valine and then eluting it using a conventional method, thus completing the present invention.
即ち、本発明方法は
イソプロピルヒダントインをアルカリ加水分解
して得られるDL−バリン水溶液からＤ−バリン
及びＬ−バリンを分離する方法において、
(1) アルカリ加水分解反応液中のDL−バリンを
アセチル化したのち酸析してＮ−アセチル−
DL−バリンを単離し、
(2) 単離したＮ−アセチル−DL−バリンにアシ
ラーゼを作用させて選択的にＮ−アセチル−Ｌ
−バリンのアセチルアミノ基のみを加水分解
し、
(3) 得られるＬ−バリンとＮ−アセチル−Ｄ−バ
リンを含む反応液のPHを１以下にしてＮ−アセ
チル−Ｄ−バリンを晶出させるとともにＬ−バ
リンを完溶させて固液分離し、
(4) Ｎ−アセチル−Ｄ−バリンを含む固相分を酸
加水分解してＤ−バリンとなしたのち、これを
晶出、単離し、
(5) Ｌ−バリンを含む酸性濾液は強酸性陽イオン
交換樹脂と接触させてＬ−バリンを吸着後溶離
し、さらに単離する。 That is, the method of the present invention is a method for separating D-valine and L-valine from a DL-valine aqueous solution obtained by alkaline hydrolysis of isopropylhydantoin, which includes: (1) acetylating DL-valine in the alkaline hydrolysis reaction solution; After that, N-acetyl-
DL-valine is isolated, and (2) acylase is applied to the isolated N-acetyl-DL-valine to selectively convert N-acetyl-L-valine into N-acetyl-L-valine.
- Hydrolyzing only the acetylamino group of valine, (3) Bringing the pH of the resulting reaction solution containing L-valine and N-acetyl-D-valine to 1 or less to crystallize N-acetyl-D-valine. (4) The solid phase containing N-acetyl-D-valine is acid-hydrolyzed to form D-valine, which is then crystallized and isolated. (5) The acidic filtrate containing L-valine is brought into contact with a strongly acidic cation exchange resin to adsorb and elute L-valine for further isolation.
ことを特徴とするイソプロピルヒダントインから
Ｄ−バリン及びＬ−バリンの製造方法である。This is a method for producing D-valine and L-valine from isopropylhydantoin.
本発明は以下のようにして実施する。 The present invention is carried out as follows.
イソプロピルヒダントインの加水分解は、常法
にしたがい水酸化ナトリウムなどアルカリをイソ
プロピルヒダントインに対して１〜５倍、好まし
くは２〜４倍モル比、及び水を加えて、100℃以
上で実施する。イソプロピルヒダントインより
DL−バリンへの転換率は通常95％以上である。
反応を加圧下高温で行えば反応時間が短縮できる
ので好ましい。 Hydrolysis of isopropylhydantoin is carried out at 100° C. or higher by adding an alkali such as sodium hydroxide in a molar ratio of 1 to 5 times, preferably 2 to 4 times, relative to isopropylhydantoin, and water according to a conventional method. From isopropylhydantoin
The conversion rate to DL-valine is usually 95% or more.
It is preferable to carry out the reaction under pressure and at high temperature because the reaction time can be shortened.
得られるDL−バリンは単離することなく、加
水分解反応液に直接無水酢酸を添加してアセチル
化を行う。無水酢酸の添加量はDL−バリンの１
〜５倍モル、好ましくは1.5〜3.0倍モルであり、
添加温度は20〜80℃、好ましくは35〜45℃であ
る。 The obtained DL-valine is acetylated by directly adding acetic anhydride to the hydrolysis reaction solution without isolation. The amount of acetic anhydride added is 1 of DL-valine.
~5 times the molar amount, preferably 1.5 to 3.0 times the molar amount,
The addition temperature is 20-80°C, preferably 35-45°C.
無水酢酸添加後数時間撹拌してDL−バリンを
Ｎ−アセチル−DL−バリンへ転換する。アセチ
ル化終了後、反応液を０〜10℃まで冷却し、強
酸、例えば35％塩酸などを加えて液を酸性側、好
ましくはPH２以下となるように調整後、析出する
Ｎ−アセチル−DL−バリンを濾別する。 After addition of acetic anhydride, the mixture is stirred for several hours to convert DL-valine to N-acetyl-DL-valine. After the acetylation is completed, the reaction solution is cooled to 0 to 10°C, and a strong acid such as 35% hydrochloric acid is added to adjust the solution to an acidic side, preferably PH2 or less, and the precipitated N-acetyl-DL- Filter off the valine.
得られるＮ−アセチル−DL−バリンは湿体の
まま、もしくは乾燥後、水に溶解してアシラーゼ
により光学分割する。Ｎ−アセチル−DL−バリ
ンの濃度は５〜40重量％、特に10〜20重量％が好
ましく、また、アルカリにより液のPHを７〜10、
好ましくは7.0〜8.0になるように調整する。さら
にアシラーゼをＮ−アセチル−DL−バリンに対
して1/400〜1/20重量部、およびCoCl2・6H2Oを
系内濃度が10-6M〜10-2Mとなるように添加し、
30〜60℃で10〜70時間かきまぜる。光学分割反応
が進行するに従つて系内にはＬ−バリンの結晶が
析出してくる。 The obtained N-acetyl-DL-valine is left wet or after drying, dissolved in water and optically resolved with acylase. The concentration of N-acetyl-DL-valine is preferably 5 to 40% by weight, particularly 10 to 20% by weight, and the pH of the liquid is adjusted to 7 to 10 with an alkali.
Adjust preferably to 7.0 to 8.0. Furthermore, 1/400 to 1/20 parts by weight of acylase to N-acetyl-DL-valine and CoCl 2 6H 2 O were added to the system concentration to 10 -6 M to 10 -2 M. ,
Stir at 30-60℃ for 10-70 hours. As the optical resolution reaction progresses, L-valine crystals begin to precipitate within the system.
分割反応終了後、Ｌ−バリンの析出している反
応マスをそのままあるいは適当量まで濃縮して強
酸、例えば35％塩酸を添加してPHを１以下となる
まで加えれば、析出していたＬ−バリンの結晶は
PHの低下とともに溶解し、Ｎ−アセチル−Ｄ−バ
リンが結晶として析出してくるので０〜10℃で数
時間晶出を行い、常法にしたがい固液分離し、Ｎ
−アセチル−Ｄ−バリンの結晶を得る。 After the splitting reaction is completed, the precipitated L-valine can be removed as is or by concentrating the reaction mass to an appropriate amount and adding a strong acid, such as 35% hydrochloric acid, until the pH is below 1. valine crystals
It dissolves as the pH decreases, and N-acetyl-D-valine precipitates as crystals, so crystallization is carried out at 0 to 10°C for several hours, followed by solid-liquid separation according to the usual method.
- Obtain crystals of acetyl-D-valine.
得られたＮ−アセチル−Ｄ−バリンは、通常純
度98％以上、分割反応に仕込んだＮ−アセチル−
DL−バリンに対して45％以上の収率であり、光
学純度90％以上のものが得られる。 The obtained N-acetyl-D-valine usually has a purity of 98% or more, and the N-acetyl-D-valine charged in the separation reaction has a purity of 98% or higher.
The yield is 45% or more based on DL-valine, and the optical purity is 90% or more.
このようにして得られたＮ−アセチル−Ｄ−バ
リンに水および塩酸を加え、加熱還流して加水分
解を行う。使用する塩酸の量はＮ−アセチル−Ｄ
−バリンに対し１〜３倍モル、特に好ましいのは
1.5倍モル程度であり、反応時のＮ−アセチル−
Ｄ−バリンの濃度は10〜40重量％、更に好ましく
は25〜35重量％である。反応は数時間、好ましく
は５時間以上続けるのがよい。 Water and hydrochloric acid are added to the N-acetyl-D-valine thus obtained, and the mixture is heated under reflux to perform hydrolysis. The amount of hydrochloric acid used is N-acetyl-D
- 1 to 3 times the mole relative to valine, particularly preferably
It is about 1.5 times the molar amount, and N-acetyl-
The concentration of D-valine is 10 to 40% by weight, more preferably 25 to 35% by weight. The reaction is preferably continued for several hours, preferably for 5 hours or more.
加水分解終了後、水酸化ナトリウムのようなア
ルカリを加えてPH５〜７に調整し、０〜10℃に冷
却する。同温度に数時間保つたのち、析出する結
晶を濾過、乾燥してＤ−バリンを得る。こうして
得られるＤ−バリンの純度、光学純度ともに99％
以上である。 After the hydrolysis is completed, an alkali such as sodium hydroxide is added to adjust the pH to 5 to 7, and the mixture is cooled to 0 to 10°C. After keeping at the same temperature for several hours, the precipitated crystals are filtered and dried to obtain D-valine. The purity and optical purity of D-valine obtained in this way are both 99%.
That's all.
一方、Ｎ−アセチル−Ｄ−バリンを濾別したあ
との濾液中には通常、Ｌ−バリン約10％、Ｎ−ア
セチルバリン約３％（Ｄ−体、Ｌ−体合計）、塩
化ナトリウム約10％、塩酸約３％程度が含有され
ている。これを強酸性陽イオン交換樹脂と接触さ
せてＬ−バリンを吸着させるのであるが、その際
イオン交換樹脂は塔に充填し、それに濾液を通液
するのが望ましい。また、使用するイオン交換樹
脂量は、その交換容量が、通液する濾液中のＬ−
バリンと塩化ナトリウムの合計等量以上、好まし
くは1.5〜2.0倍等量になるような量にするのが適
当である。 On the other hand, the filtrate after filtering off N-acetyl-D-valine usually contains about 10% L-valine, about 3% N-acetylvaline (total of D-form and L-form), and about 10% sodium chloride. %, and about 3% hydrochloric acid. This is brought into contact with a strongly acidic cation exchange resin to adsorb L-valine. In this case, it is desirable to fill a column with the ion exchange resin and pass the filtrate through it. In addition, the amount of ion exchange resin used is such that its exchange capacity is L-
It is appropriate to use an amount that is equal to or more than the total amount of valine and sodium chloride, preferably 1.5 to 2.0 times the equivalent amount.
強酸性イオン交換樹脂によるＬ−バリンの吸着
に際しては、同時に含有している塩化ナトリウム
のナトリウムイオンも吸着されるが、これは充填
塔での吸着ゾーンがＬ−バリンとは異なるので、
溶離において容易に分離でき、また好ましい本発
明方法として、複数の充填塔を直列に連結してお
き通液すれば、最初の塔でナトリウムイオンのみ
が吸着され除去できるので分離が容易である。 When adsorbing L-valine with a strongly acidic ion exchange resin, the sodium ions of sodium chloride contained therein are also adsorbed at the same time, but this is because the adsorption zone in the packed column is different from that of L-valine.
They can be easily separated by elution, and as a preferred method of the present invention, if a plurality of packed columns are connected in series and the liquid is passed through them, only sodium ions can be adsorbed and removed in the first column, making separation easy.
また、濾液中のＮ−アセチルバリンおよび塩酸
はイオン交換樹脂には吸着されずにそのまま排出
されるので、必要ならば排出液からＮ−アセチル
バリンを回収、循環使用することもできる。 Furthermore, since N-acetylvaline and hydrochloric acid in the filtrate are discharged as they are without being adsorbed by the ion exchange resin, N-acetylvaline can be recovered from the discharged solution and recycled if necessary.
Ｌ−バリンを吸着したイオン交換樹脂に適当な
溶離剤、例えばアンモニア水を通液すると、Ｌ−
バリンは溶離されるので適当量まで濃縮後、冷却
し、析出する結晶を濾過、乾燥すればＬ−バリン
が得られる。この晶出操作の際、低級アルコール
（メタノール、エタノール、ｎ−プロパノール、
イソプロパノールなど）を添加することも収率、
品質向上のために有効である。 When a suitable eluent, such as aqueous ammonia, is passed through an ion exchange resin that has adsorbed L-valine, L-valine is released.
Since valine is eluted, L-valine can be obtained by concentrating to an appropriate amount, cooling, filtering and drying the precipitated crystals. During this crystallization operation, lower alcohols (methanol, ethanol, n-propanol,
The addition of isopropanol (such as isopropanol) also improves the yield,
Effective for improving quality.
こうして得られるＬ−バリンの純度、光学純度
はいずれも99％以上である。 The purity and optical purity of L-valine thus obtained are both 99% or higher.
実施例 以下、実施例にて詳しく説明する。Example This will be explained in detail in Examples below.
実施例 １
イソプロピルヒダントイン416.6ｇ（2.89モル）
に水468.5ｇおよび45％水酸化ナトリウム水溶液
781.3ｇ（8.79モル）を加え、常圧下、かきまぜ
ながら24時間加熱還流した。得られた加水分解反
応液に、強くかきまぜつつ無水酢酸448ｇを40℃、
２時間で滴下し、さらに同温度に４時間保温し
た。ついで０〜10℃に冷却したのち、35％塩酸
600ｇを約２時間かけて滴下してPHを２とし、さ
らに０〜10℃に約４時間保つてＮ−アセチル−
DL−バリンの結晶を析出させた。これを濾過し
てＮ−アセチル−DL−バリンの湿体920ｇ（乾燥
後の重量は431.3ｇ）を得た。Example 1 Isopropylhydantoin 416.6g (2.89mol)
468.5 g of water and 45% aqueous sodium hydroxide solution
781.3 g (8.79 mol) was added thereto, and the mixture was heated under reflux for 24 hours with stirring under normal pressure. Add 448 g of acetic anhydride to the resulting hydrolysis reaction solution at 40°C while stirring vigorously.
The mixture was added dropwise over 2 hours and kept at the same temperature for an additional 4 hours. Then, after cooling to 0-10℃, add 35% hydrochloric acid.
600g was added dropwise over about 2 hours to adjust the pH to 2, and then kept at 0 to 10℃ for about 4 hours to obtain N-acetyl-
Crystals of DL-valine were precipitated. This was filtered to obtain 920 g of wet N-acetyl-DL-valine (weight after drying: 431.3 g).
得られたＮ−アセチル−DL−バリンの湿体全
量に水258ｇおよび45％水酸化ナトリウム水溶液
250ｇを加えて溶解し（PHは7.5となる）、さらに
天野製薬社製アシラーゼ7.8ｇとCoCl2・6H2O2.1
ｇを添加して40℃で４時間かきまぜ、Ｎ−アセチ
ル−Ｌ−バリンのみを選択的に加水分解した。 Add 258 g of water and 45% aqueous sodium hydroxide solution to the entire wet amount of N-acetyl-DL-valine obtained.
Add 250g and dissolve (PH becomes 7.5), then add 7.8g of Amano Pharmaceutical's acylase and CoCl 2 6H 2 O2.1
g was added and stirred at 40°C for 4 hours to selectively hydrolyze only N-acetyl-L-valine.
次いで40℃に保ちながら35％塩酸380ｇを加え
てPHを1.0としたのち冷却し、０〜10℃に２時間
保つた。析出した結晶を濾過、水洗（100ml）す
ることにより、Ｎ−アセチル−Ｄ−バリンの湿体
242.6ｇ、濾洗液1675ｇを得、湿体を乾燥してＮ
−アセチル−Ｄ−バリン194.1ｇを得た。このも
のの純度は98.8％、光学純度は94.2％、〔α〕D 20＋
16.8°（Ｃ＝１、水）であつた。 Next, while maintaining the temperature at 40°C, 380 g of 35% hydrochloric acid was added to adjust the pH to 1.0, and the mixture was cooled and kept at 0 to 10°C for 2 hours. By filtering the precipitated crystals and washing with water (100 ml), wet N-acetyl-D-valine was obtained.
242.6g and 1675g of filtration and washing liquid were obtained, and the wet material was dried and N
194.1 g of -acetyl-D-valine was obtained. The purity of this product is 98.8%, the optical purity is 94.2%, [α] D 20 +
It was 16.8° (C=1, water).
こうして得られたＮ−アセチル−Ｄ−バリンの
全量を水497ｇ、35％塩酸150ｇと混合し、５時間
加熱還流してＤ−バリンの酸性水溶液を得た。こ
れに45％水酸化ナトリウム水溶液を40℃で滴下し
てPHを5.5に調整後、冷却し、０〜10℃に２時間
保つた。析出した結晶を濾過、乾燥することによ
りＤ−バリン115.0ｇ（0.981モル）を得た。収率
は33.9％（対イソプロピルヒダントイン）であ
り、本品の純度は99.6％、〔α〕D 20−27.5°（Ｃ＝８
、
6NHCl）であつた、
一方、強酸性陽イオン交換樹脂レバチツトＳ−
100（バイエル社製）2.0を充填した第１塔と、
0.8充填した第２塔を直列に連結した樹脂塔に、
Ｎ−アセチル−Ｄ−バリンを濾別した際に得られ
た濾洗液1675ｇ（Ｌ−バリン8.5％、Ｎ−アセチ
ルバリン2.6％、塩化ナトリウム約10％、塩酸約
３％含有）を下向流で通液した（SV5）。 The total amount of N-acetyl-D-valine thus obtained was mixed with 497 g of water and 150 g of 35% hydrochloric acid, and the mixture was heated under reflux for 5 hours to obtain an acidic aqueous solution of D-valine. A 45% aqueous sodium hydroxide solution was added dropwise thereto at 40°C to adjust the pH to 5.5, then cooled and kept at 0 to 10°C for 2 hours. The precipitated crystals were filtered and dried to obtain 115.0 g (0.981 mol) of D-valine. The yield was 33.9% (based on isopropylhydantoin), the purity of this product was 99.6%, [α] D 20 -27.5° (C = 8
,
6NHCl), on the other hand, the strongly acidic cation exchange resin Revacit S-
A first column filled with 100 (manufactured by Bayer) 2.0,
The resin tower is connected in series with the second tower filled with 0.8
1675 g of the filtrate and washing liquid obtained when N-acetyl-D-valine was filtered out (containing 8.5% L-valine, 2.6% N-acetylvaline, about 10% sodium chloride, and about 3% hydrochloric acid) was passed through a downward flow. (SV5).
この操作により濾洗液中のナトリウムイオンは
第１塔に、Ｌ−バリンは第２塔に吸着され、塩酸
およびＮ−アセチルバリンは吸着されずに塔外へ
流下した。 As a result of this operation, sodium ions in the filtrate and washing liquid were adsorbed in the first column, L-valine was adsorbed in the second column, and hydrochloric acid and N-acetylvaline were not adsorbed and flowed out of the column.
次いで、第２塔に純水３を通液（SV5）して
洗浄後、５％アンモニア水１を通液（SV5）し
て吸着したＬ−バリンを溶離した。Ｌ−バリンを
含む溶離液１を約半量になるまで濃縮し、35％
塩酸を加えてPH6.5に調整後、イソプロパノール
1.0を加えて０〜10℃に冷却した。同温度に２
時間保つたのち、析出した結晶を濾過、乾燥して
Ｌ−バリン110.0ｇ（0.939モル）を得た。収率は
32.5％（対イソプロピルヒダントイン）であり、
本品の純度は99.3％、〔α〕D 20＋27.2°（Ｃ＝８、
6NHCl）であつた。 Next, 3 parts of pure water were passed through the second column (SV5) for washing, and 1 part of 5% ammonia water was passed therethrough (SV5) to elute the adsorbed L-valine. Concentrate eluent 1 containing L-valine to about half its volume to 35%.
After adjusting the pH to 6.5 by adding hydrochloric acid, add isopropanol.
1.0 and cooled to 0-10°C. 2 at the same temperature
After holding for a certain period of time, the precipitated crystals were filtered and dried to obtain 110.0 g (0.939 mol) of L-valine. The yield is
32.5% (vs. isopropylhydantoin),
The purity of this product is 99.3%, [α] D 20 +27.2° (C = 8,
6NHCl).
イソプロピルヒダントインに対するＤ−バリン
とＬ−バリンの収率を合計すると66.4％に達し
た。 The total yield of D-valine and L-valine based on isopropylhydantoin reached 66.4%.
Claims (1)
分解して得られるDL−バリン水溶液からＤ−バ
リン及びＬ−バリンを分離する方法において、 (1) アルカリ加水分解反応液中のDL−バリンを
アセチル化したのち酸析してＮ−アセチル−
DL−バリンを単離し、 (2) 単離したＮ−アセチル−DL−バリンにアシ
ラーゼを作用させて選択的にＮ−アセチル−Ｌ
−バリンのアセチルアミノ基のみを加水分解
し、 (3) 得られるＬ−バリンとＮ−アセチル−Ｄ−バ
リンを含む反応液のPHを１以下にしてＮ−アセ
チル−Ｄ−バリンを晶出させるとともにＬ−バ
リンを完溶させて固液分離し、 (4) Ｎ−アセチル−Ｄ−バリンを含む固相分を、
酸加水分解してＤ−バリンとなしたのち、これ
を晶出、単離し、 (5) Ｌ−バリンを含む酸性濾液は、強酸性陽イオ
ン交換樹脂と接触させてＬ−バリンを吸着後、
溶離し、さらに単離する ことを特徴とするイソプロピルヒダントインから
Ｄ−バリン及びＬ−バリンの製造方法。[Scope of Claims] 1. A method for separating D-valine and L-valine from a DL-valine aqueous solution obtained by alkaline hydrolysis of isopropylhydantoin, comprising: (1) DL-valine in an alkaline hydrolysis reaction solution; After acetylation, acid precipitation produces N-acetyl-
DL-valine is isolated, and (2) acylase is applied to the isolated N-acetyl-DL-valine to selectively convert N-acetyl-L into
- Hydrolyzing only the acetylamino group of valine, (3) Bringing the pH of the resulting reaction solution containing L-valine and N-acetyl-D-valine to 1 or less to crystallize N-acetyl-D-valine. (4) The solid phase containing N-acetyl-D-valine is
After acid hydrolysis to form D-valine, this is crystallized and isolated. (5) The acidic filtrate containing L-valine is brought into contact with a strongly acidic cation exchange resin to adsorb L-valine.
A method for producing D-valine and L-valine from isopropylhydantoin, which comprises elution and further isolation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP18653084A JPS6167498A (en) | 1984-09-07 | 1984-09-07 | Production of d-valine and l-valine from isopropyl hydantoin |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP18653084A JPS6167498A (en) | 1984-09-07 | 1984-09-07 | Production of d-valine and l-valine from isopropyl hydantoin |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS6167498A JPS6167498A (en) | 1986-04-07 |
| JPH0338840B2 true JPH0338840B2 (en) | 1991-06-11 |
Family
ID=16190104
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP18653084A Granted JPS6167498A (en) | 1984-09-07 | 1984-09-07 | Production of d-valine and l-valine from isopropyl hydantoin |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS6167498A (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS632960A (en) * | 1986-06-20 | 1988-01-07 | Showa Denko Kk | Production of valine |
-
1984
- 1984-09-07 JP JP18653084A patent/JPS6167498A/en active Granted
Also Published As
| Publication number | Publication date |
|---|---|
| JPS6167498A (en) | 1986-04-07 |
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