JPS6225359B2 - - Google Patents
Info
- Publication number
- JPS6225359B2 JPS6225359B2 JP53081954A JP8195478A JPS6225359B2 JP S6225359 B2 JPS6225359 B2 JP S6225359B2 JP 53081954 A JP53081954 A JP 53081954A JP 8195478 A JP8195478 A JP 8195478A JP S6225359 B2 JPS6225359 B2 JP S6225359B2
- Authority
- JP
- Japan
- Prior art keywords
- phenylglycine
- hydroxy
- hydroxymethyl
- aminoacylase
- acetyl
- 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
- 108010003977 aminoacylase I Proteins 0.000 claims description 14
- ULGJWNIHLSLQPZ-UHFFFAOYSA-N 7-[(6,8-dichloro-1,2,3,4-tetrahydroacridin-9-yl)amino]-n-[2-(1h-indol-3-yl)ethyl]heptanamide Chemical compound C1CCCC2=NC3=CC(Cl)=CC(Cl)=C3C(NCCCCCCC(=O)NCCC=3C4=CC=CC=C4NC=3)=C21 ULGJWNIHLSLQPZ-UHFFFAOYSA-N 0.000 claims description 12
- 238000006460 hydrolysis reaction Methods 0.000 claims description 10
- 238000004519 manufacturing process Methods 0.000 claims description 6
- 230000000813 microbial effect Effects 0.000 claims description 6
- 230000002538 fungal effect Effects 0.000 claims description 5
- 230000007062 hydrolysis Effects 0.000 claims description 5
- 241000589151 Azotobacter Species 0.000 claims description 4
- ZGYNTYWHXNWAFO-UHFFFAOYSA-N 2-[3-hydroxy-4-(hydroxymethyl)anilino]acetic acid Chemical compound OCC1=CC=C(NCC(O)=O)C=C1O ZGYNTYWHXNWAFO-UHFFFAOYSA-N 0.000 claims description 2
- IVQDNDSBHMVGLM-UHFFFAOYSA-N 2-amino-2-[4-(hydroxymethyl)phenyl]acetic acid Chemical compound OC(=O)C(N)C1=CC=C(CO)C=C1 IVQDNDSBHMVGLM-UHFFFAOYSA-N 0.000 claims 1
- ZGUNAGUHMKGQNY-SSDOTTSWSA-N D-alpha-phenylglycine Chemical compound OC(=O)[C@H](N)C1=CC=CC=C1 ZGUNAGUHMKGQNY-SSDOTTSWSA-N 0.000 claims 1
- 125000002777 acetyl group Chemical group [H]C([H])([H])C(*)=O 0.000 claims 1
- 125000003236 benzoyl group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C(*)=O 0.000 claims 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 10
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 9
- 150000001413 amino acids Chemical class 0.000 description 7
- 239000000243 solution Substances 0.000 description 7
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 5
- 238000006243 chemical reaction Methods 0.000 description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 5
- 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 description 4
- NKFISLRONBSNFI-UHFFFAOYSA-N 2-benzamido-2-[3-hydroxy-4-(hydroxymethyl)phenyl]acetic acid Chemical compound C(C1=CC=CC=C1)(=O)NC(C1=CC(=C(C=C1)CO)O)C(=O)O NKFISLRONBSNFI-UHFFFAOYSA-N 0.000 description 4
- 239000003456 ion exchange resin Substances 0.000 description 4
- 229920003303 ion-exchange polymer Polymers 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 108700023418 Amidases Proteins 0.000 description 3
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 3
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 3
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 3
- 230000002378 acidificating effect Effects 0.000 description 3
- 102000005922 amidase Human genes 0.000 description 3
- -1 benzoyl-3-hydroxy-4-(hydroxymethyl)-D-phenylglycine Chemical compound 0.000 description 3
- 239000003610 charcoal Substances 0.000 description 3
- 239000000843 powder Substances 0.000 description 3
- TWGMQVBSQMTMCB-SNVBAGLBSA-N (2R)-2-acetamido-2-[3-hydroxy-4-(hydroxymethyl)phenyl]acetic acid Chemical compound C(C)(=O)N[C@H](C1=CC(=C(C=C1)CO)O)C(=O)O TWGMQVBSQMTMCB-SNVBAGLBSA-N 0.000 description 2
- TWGMQVBSQMTMCB-UHFFFAOYSA-N 2-acetamido-2-[3-hydroxy-4-(hydroxymethyl)phenyl]acetic acid Chemical compound CC(=O)NC(C(O)=O)C1=CC=C(CO)C(O)=C1 TWGMQVBSQMTMCB-UHFFFAOYSA-N 0.000 description 2
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 description 2
- NTIZESTWPVYFNL-UHFFFAOYSA-N Methyl isobutyl ketone Chemical compound CC(C)CC(C)=O NTIZESTWPVYFNL-UHFFFAOYSA-N 0.000 description 2
- UIHCLUNTQKBZGK-UHFFFAOYSA-N Methyl isobutyl ketone Natural products CCC(C)C(C)=O UIHCLUNTQKBZGK-UHFFFAOYSA-N 0.000 description 2
- WPYMKLBDIGXBTP-UHFFFAOYSA-N benzoic acid Chemical compound OC(=O)C1=CC=CC=C1 WPYMKLBDIGXBTP-UHFFFAOYSA-N 0.000 description 2
- 239000013078 crystal Substances 0.000 description 2
- 229920001467 poly(styrenesulfonates) Polymers 0.000 description 2
- 239000011347 resin Substances 0.000 description 2
- 229920005989 resin Polymers 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- DQLYTFPAEVJTFM-UHFFFAOYSA-N 2-azaniumyl-2-(3-hydroxyphenyl)acetate Chemical compound OC(=O)C(N)C1=CC=CC(O)=C1 DQLYTFPAEVJTFM-UHFFFAOYSA-N 0.000 description 1
- 241000228212 Aspergillus Species 0.000 description 1
- 239000005711 Benzoic acid Substances 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- ZGUNAGUHMKGQNY-ZETCQYMHSA-N L-alpha-phenylglycine zwitterion Chemical compound OC(=O)[C@@H](N)C1=CC=CC=C1 ZGUNAGUHMKGQNY-ZETCQYMHSA-N 0.000 description 1
- 150000008575 L-amino acids Chemical class 0.000 description 1
- 241000228143 Penicillium Species 0.000 description 1
- 241000187747 Streptomyces Species 0.000 description 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-N ammonia Natural products N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 235000010233 benzoic acid Nutrition 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 229920001429 chelating resin Polymers 0.000 description 1
- GFHNAMRJFCEERV-UHFFFAOYSA-L cobalt chloride hexahydrate Chemical compound O.O.O.O.O.O.[Cl-].[Cl-].[Co+2] GFHNAMRJFCEERV-UHFFFAOYSA-L 0.000 description 1
- GVPFVAHMJGGAJG-UHFFFAOYSA-L cobalt dichloride Chemical compound [Cl-].[Cl-].[Co+2] GVPFVAHMJGGAJG-UHFFFAOYSA-L 0.000 description 1
- 239000013065 commercial product Substances 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 238000000921 elemental analysis Methods 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 125000004029 hydroxymethyl group Chemical group [H]OC([H])([H])* 0.000 description 1
- 229960001438 immunostimulant agent Drugs 0.000 description 1
- 239000003022 immunostimulating agent Substances 0.000 description 1
- 230000003308 immunostimulating effect Effects 0.000 description 1
- 239000000543 intermediate Substances 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 239000012466 permeate Substances 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000001953 recrystallisation Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/52—Improvements relating to the production of bulk chemicals using catalysts, e.g. selective catalysts
Landscapes
- Enzymes And Modification Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Description
【発明の詳細な説明】
本発明は光学活性３−ヒドロキシ−４−（ヒド
ロキシメチル）フエニルグリシンの製造法に関す
るものである。さらに詳しくは、Ｎアセチル−も
しくはＮ−ベンゾイル−３−ヒドロキシ−４−
（ヒドロキシメチル）−DL−フエニルグリシンに
微生物起原のアミノアシラーゼを作用させて、３
−ヒドロキシ−４−（ヒドロキシメチル）−Ｌ−フ
エニルグリシンおよびＮ−アセチル−もしくはＮ
−ベンゾイル−３−ヒドロキシ−４−（ヒドロキ
シメチル）−Ｄ−フエニルグリシンに不斉加水分
解し、両者を分離採取することからなる光学活性
３−ヒドロキシ−４−（ヒドロキシメチル）フエ
ニルグリシンの製造法に関するものである。
本発明の出発物質であるＮ−アセチル−もしく
はＮ−ベンゾイル−３−ヒドロキシ−４−（ヒド
ロキシメチル）−DL−フエニルグリシンは、たと
えば、Ｎ−アセチル−もしくはＮ−ベンゾイル−
３−ヒドロキシ−DL−フエニルグリシンに塩基
の存在下にホルムアルデヒドを作用させることに
よつて容易に製造することができる。
これらのＮ−アシルアミノ酸の不斉加水分解は
糸状菌アミノアシラーゼもしくはアゾトバクター
属菌株の産生するアシラーゼなどの微生物起原の
アミノアシラーゼによつて容易に行なわれる。糸
状菌アミノアシラーゼはたとえばアスペルギルス
属あるいはペニシリウム属などの糸状菌の培養物
からその産生するアミノアシラーゼを採取して得
られ、市販品として入手することができる。また
アミノアシラーゼを産生するアゾトバクター属菌
体としてはたとえばNo.1736をあげることができ
る。
その菌学的性状は特公昭45−8634に記載されて
いる。
微生物起原のアミノアシラーゼによる不斉加水
分解反応は、たとえばPH６−８、温度約35−40℃
で行なわれ、反応の進行状況はたとえば高速液体
クロマトグラフイーによつて反応液中のＮ−アシ
ルアミノ酸と遊離アミノ酸の比率を追跡すること
によつて知ることができる。
不斉加水分解反応液から３−ヒドロキシ−４−
（ヒドロキシメチル）−Ｌ−フエニルグリシンおよ
びＮ−アセチル−もしくはＮ−ベンゾイル−３−
ヒドロキシ−４−（ヒドロキシメチル）−Ｄ−フエ
ニルグリシンを分離採取するためには、たとえば
強酸性イオン交換樹脂によつて遊離アミノ酸のみ
を吸着させる方法が有利である。吸着した遊離ア
ミノ酸はアンモニア水によつて溶出され、この溶
出液を減圧濃縮すれば遊離のＬ−アミノ酸が析出
する。またイオン交換樹脂塔の通過液からＮ−ア
シル−Ｄ−アミノ酸が回収される。
３−ヒドロキシ−４−（ヒドロキシメチル）−Ｌ
−フエニルグリシンはたとえば免疫賦活剤として
有用である。その製法としては、従来、ストレプ
トミセス属菌株の生産する４−ホルミル−３−ヒ
ドロキシ−Ｌ−フエニルグリシンを還元する方法
が知られているが、本発明の方法によつて、化学
的合成ルートによるDL−型の中間体を光学分割
することができるようになつたものである。
以下に実施例をあげて本発明を具体的に説明す
る。
実施例 １
Ｎ−アセチル−３−ヒドロキシ−４−（ヒドロ
キシメチル）−DL−フエニルグリシン3.0ｇを
0.5N水酸化ナトリウムに溶解する。PH7.5に調整
したのち全液量を60mlとし、これに塩化コバルト
６水塩８mgおよび糸状菌アミノアシラーゼ（天野
製薬製アシラーゼアマノ、約15000単位／ｇ）100
mgを加えて、37℃で48時間不斉加水分解反応を行
なう。
反応液を炭末脱色し、強酸性イオン交換樹脂ダ
ウエツクス50W−X4（H+）30mlを充填した塔に
通液して、不斉加水分解によつて生成したアミノ
酸を吸着させる。通過液を約50mlまで減圧濃縮し
て、ダイヤイオンHP−20のカラムで分画する
と、Ｎ−アセチル−３−ヒドロキシ−４−（ヒド
ロキシメチル）−Ｄ−フエニルグリシン1.5ｇが得
られる。水から再結晶して精製することができ
る。
m.p.172℃（分解）。〔α〕２０ Ｄ−206゜（ｃ1.0、
EtOH）。
IR（KBr）νmax（cm-1）：3440、3365、3215、
2905、1715、1610、1580、1530、1430、1370、
1340、1295、1250、1220、1155、1110、1025、
960、910、865、820、730、
またアミノ酸を吸着した樹脂塔を水洗後、Ｎ
NH4OH250mlにて溶出し、溶出液を減圧濃縮して
３−ヒドロキシ−４−（ヒドロキシメチル）−Ｌ−
フエニルグリシン1.09ｇが得られた。m.p.223℃
（分解）。〔α〕２０ Ｄ131゜（ｃ1.0、Ｎ HCl）。
C9H11NO4としての元素分析値（％）
Ｃ Ｈ Ｎ
計算値 54.82 5.62 7.10
実験値 54.69 5.69 7.06
実施例 ２
Ｎ−アセチル−３−ヒドロキシ−４−（ヒドロ
キシメチル）−DL−フエニルグリシン200ｇを
0.5N水酸化ナトリウムに溶解してPH7.5に調整
し、全液量を4Lとしたのち、糸状菌アミノアシ
ラーゼ（天野製薬製アシラーゼアマノ、約15000
単位／ｇ）５ｇおよび塩化コバルト0.4ｇを加え
て、37℃で24時間不斉加水分解反応を行なう。
反応液を炭末脱色し、強酸性イオン交換樹脂ダ
ウエツクス50W−X4（H+）1.33Lを充填した塔に
通液して、不斉加水分解によつて生成したアミノ
酸を吸着させる。このカラムを水洗後、Ｎ
NH4OHで溶出し、溶出液3.5Lを集めて減圧濃縮
すると、３−ヒドロキシ−４−（ヒドロキシメチ
ル）−Ｌ−フエニルグリシン76.3ｇ（収率92.5
％）が得られる。さらに精製するために、この製
品158ｇをアンモニア水溶液に溶解して炭末脱色
し、約800mlまで減圧濃縮して一夜冷却し、析出
晶を冷水およびエタノールで洗浄して乾燥する。
収量146ｇ。〔α〕２４ Ｄ130゜（ｃ1.0、Ｎ HCl）。
また樹脂塔通過液6Lを約300mlまで減圧濃縮し
て冷却すると、Ｎ−アセチル−３−ヒドロキシ−
４−（ヒドロキシメチル）−Ｄ−フエニルグリシン
が析出する。収量95.2ｇ。〔α〕２４ Ｄ−193゜（ｃ
1.0、EtOH）。
実施例 ３
Ｎ−ベンゾイル−３−ヒドロキシ−４−（ヒド
ロキシメチル）−DL−フエニルグリシン16.5ｇを
水酸化ナトリウム水溶液に溶解してPH8.0に調整
したのち全液量を800mlとし、これにアゾトバク
ター属菌株No.1736のアセトン乾燥菌体1.6ｇを加
えて、34−36℃で24時間不斉加水分解反応を行な
う。
反応液を塩酸でPH2.0に調整してメチルイソ
ブチルケトンで抽出後、水層をアンバーライト
IR120を充填した塔に通液してアミノ酸を吸着
し、ついで3N NH4OHで溶出する。溶出液を減
圧濃縮して析出する結晶を75％含水エタノールで
洗浄して、３−ヒドロキシ−４−（ヒドロキシメ
チル）−Ｌ−フエニルグリシンが得られた。収量
2.68ｇ。〔α〕２４ Ｄ130゜（ｃ1.0、Ｎ HCl）。
また、メチルイソブチルケトン抽出液をPH8.0
にて水で再抽出し、水層をPH2.0に調整してから
ベンゼンで抽出して副生安息香酸を除去する。さ
らに酢酸エチルで抽出し、抽出液を減圧濃縮する
とＮ−ベンゾイル−３−ヒドロキシ−４−（ヒド
ロキシメチル）−Ｄ−フエニルグリシンが析出す
る。収量8.2ｇ。
〔α〕２４ Ｄ−72゜（ｃ１、EtOH）。 DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method for producing optically active 3-hydroxy-4-(hydroxymethyl)phenylglycine. More specifically, N-acetyl- or N-benzoyl-3-hydroxy-4-
(Hydroxymethyl)-DL-phenylglycine is treated with aminoacylase of microbial origin, and 3
-Hydroxy-4-(hydroxymethyl)-L-phenylglycine and N-acetyl- or N
-Asymmetric hydrolysis to benzoyl-3-hydroxy-4-(hydroxymethyl)-D-phenylglycine and separation and collection of both. It concerns the manufacturing method. The starting material according to the invention, N-acetyl- or N-benzoyl-3-hydroxy-4-(hydroxymethyl)-DL-phenylglycine, is, for example, N-acetyl- or N-benzoyl-3-hydroxy-4-(hydroxymethyl)-DL-phenylglycine.
It can be easily produced by reacting formaldehyde with 3-hydroxy-DL-phenylglycine in the presence of a base. Asymmetric hydrolysis of these N-acyl amino acids is easily carried out by aminoacylases of microbial origin, such as filamentous fungal aminoacylases or acylases produced by Azotobacter strains. Filamentous fungal aminoacylase is obtained by collecting the aminoacylase produced by a culture of filamentous fungi such as Aspergillus or Penicillium, and can be obtained as a commercial product. An example of an Azotobacter cell that produces aminoacylase is No. 1736. Its mycological properties are described in Japanese Patent Publication No. 45-8634. The asymmetric hydrolysis reaction by aminoacylase of microbial origin is performed at a pH of 6-8 and a temperature of approximately 35-40°C.
The progress of the reaction can be determined by monitoring the ratio of N-acyl amino acids to free amino acids in the reaction solution using, for example, high performance liquid chromatography. 3-hydroxy-4- from the asymmetric hydrolysis reaction solution
(Hydroxymethyl)-L-phenylglycine and N-acetyl- or N-benzoyl-3-
In order to separate and collect hydroxy-4-(hydroxymethyl)-D-phenylglycine, it is advantageous to adsorb only free amino acids using, for example, a strongly acidic ion exchange resin. The adsorbed free amino acids are eluted with aqueous ammonia, and when this eluate is concentrated under reduced pressure, free L-amino acids are precipitated. Further, N-acyl-D-amino acids are recovered from the liquid passing through the ion exchange resin tower. 3-hydroxy-4-(hydroxymethyl)-L
- Phenylglycine is useful, for example, as an immunostimulant. Conventionally, the production method is known to reduce 4-formyl-3-hydroxy-L-phenylglycine produced by Streptomyces strains, but the method of the present invention allows for chemical synthesis route. It has now become possible to optically resolve DL-type intermediates. The present invention will be specifically explained below with reference to Examples. Example 1 3.0 g of N-acetyl-3-hydroxy-4-(hydroxymethyl)-DL-phenylglycine
Dissolve in 0.5N sodium hydroxide. After adjusting the pH to 7.5, the total volume of the liquid was reduced to 60 ml, and to this was added 8 mg of cobalt chloride hexahydrate and 100 mg of filamentous fungal aminoacylase (Acylase Amano manufactured by Amano Pharmaceutical Co., Ltd., approximately 15,000 units/g).
mg was added, and the asymmetric hydrolysis reaction was carried out at 37°C for 48 hours. The reaction solution is decolorized with charcoal powder and passed through a tower filled with 30 ml of strongly acidic ion exchange resin Dowex 50W-X4 (H + ) to adsorb the amino acids produced by asymmetric hydrolysis. The permeate was concentrated under reduced pressure to about 50 ml and fractionated using a Diaion HP-20 column to obtain 1.5 g of N-acetyl-3-hydroxy-4-(hydroxymethyl)-D-phenylglycine. It can be purified by recrystallization from water. mp172℃ (decomposed). [α] 20 D -206° ( c 1.0,
EtOH). IR (KBr) νmax (cm -1 ): 3440, 3365, 3215,
2905, 1715, 1610, 1580, 1530, 1430, 1370,
1340, 1295, 1250, 1220, 1155, 1110, 1025,
960, 910, 865, 820, 730, and after washing the resin tower that adsorbed amino acids with water,
Elute with 250 ml of NH 4 OH and concentrate the eluate under reduced pressure to obtain 3-hydroxy-4-(hydroxymethyl)-L-
1.09 g of phenylglycine was obtained. mp223℃
(Disassembly). [α] 20 D 131° ( c 1.0, N HCl). Elemental analysis value (%) as C 9 H 11 NO 4 C H N Calculated value 54.82 5.62 7.10 Experimental value 54.69 5.69 7.06 Example 2 N-acetyl-3-hydroxy-4-(hydroxymethyl)-DL-phenylglycine 200g
After dissolving in 0.5N sodium hydroxide and adjusting the pH to 7.5 to bring the total volume to 4L, add filamentous fungal aminoacylase (Acylase Amano manufactured by Amano Pharmaceutical Co., Ltd., approximately 15,000
Units/g) 5g and cobalt chloride 0.4g are added, and the asymmetric hydrolysis reaction is carried out at 37°C for 24 hours. The reaction solution is decolorized with charcoal powder and passed through a tower packed with 1.33 L of strongly acidic ion exchange resin Dowex 50W-X4 (H + ) to adsorb the amino acids produced by asymmetric hydrolysis. After washing this column with water, N
Elution was performed with NH 4 OH, and 3.5 L of the eluate was collected and concentrated under reduced pressure to yield 76.3 g of 3-hydroxy-4-(hydroxymethyl)-L-phenylglycine (yield: 92.5
%) is obtained. For further purification, 158 g of this product is dissolved in an ammonia aqueous solution to decolorize the charcoal powder, concentrated under reduced pressure to about 800 ml, cooled overnight, and the precipitated crystals are washed with cold water and ethanol and dried.
Yield: 146g. [α] 24 D 130° ( c 1.0, N HCl). In addition, when 6L of the liquid passed through the resin tower was concentrated under reduced pressure to about 300ml and cooled, N-acetyl-3-hydroxy-
4-(Hydroxymethyl)-D-phenylglycine is precipitated. Yield: 95.2g. [α] 24 D -193゜( c
1.0, EtOH). Example 3 16.5 g of N-benzoyl-3-hydroxy-4-(hydroxymethyl)-DL-phenylglycine was dissolved in an aqueous sodium hydroxide solution and the pH was adjusted to 8.0, and the total liquid volume was adjusted to 800 ml. Add 1.6 g of acetone-dried bacterial cells of Azotobacter strain No. 1736, and carry out an asymmetric hydrolysis reaction at 34-36°C for 24 hours. After adjusting the reaction solution to PH2.0 with hydrochloric acid and extracting with methyl isobutyl ketone, the aqueous layer was extracted with Amberlite.
The solution is passed through a column packed with IR120 to adsorb amino acids, and then eluted with 3N NH 4 OH. The eluate was concentrated under reduced pressure and the precipitated crystals were washed with 75% aqueous ethanol to obtain 3-hydroxy-4-(hydroxymethyl)-L-phenylglycine. yield
2.68g. [α] 24 D 130° ( c 1.0, N HCl). In addition, methyl isobutyl ketone extract has a pH of 8.0.
Re-extract with water, adjust the aqueous layer to pH 2.0, and then extract with benzene to remove by-product benzoic acid. Further extraction is performed with ethyl acetate, and the extract is concentrated under reduced pressure to precipitate N-benzoyl-3-hydroxy-4-(hydroxymethyl)-D-phenylglycine. Yield: 8.2g. [α] 24 D −72° ( c 1, EtOH).
Claims (1)
す）で表わされるＮ−アシル−３−ヒドロキシ−
４−（ヒドロキシメチル）−DL−フエニルグリシ
ンに微生物起原のアミノアシラーゼを作用させて
不斉加水分解して該加水分解液から３−ヒドロキ
シ−４−（ヒドロキシメチル）−Ｌ−フエニルグリ
シンおよびＮ−アセチル−もしくはＮ−ベンゾイ
ル−３−ヒドロキシ−４−（ヒドロキシメチル）−
Ｄ−フエニルグリシンを分離採取することを特徴
とする光学活性３−ヒドロキシ−４−（ヒドロキ
シメチル）フエニルグリシンの製造法。 ２ 微生物起原のアミノアシラーゼが糸状菌アミ
ノアシラーゼである特許請求の範囲第１項記載の
製造法。 ３ 微生物起原のアミノアシラーゼがアゾトバク
ター属菌株の生産するアミノアシラーゼである特
許請求の範囲第１項記載の製造法。[Claims] 1 formula N-acyl-3-hydroxy- (wherein R represents an acetyl group or a benzoyl group)
4-(Hydroxymethyl)-DL-phenylglycine is asymmetrically hydrolyzed by the action of aminoacylase of microbial origin, and 3-hydroxy-4-(hydroxymethyl)-L-phenylglycine is produced from the hydrolysis solution. and N-acetyl- or N-benzoyl-3-hydroxy-4-(hydroxymethyl)-
A method for producing optically active 3-hydroxy-4-(hydroxymethyl)phenylglycine, which comprises separating and collecting D-phenylglycine. 2. The production method according to claim 1, wherein the aminoacylase of microbial origin is a filamentous fungal aminoacylase. 3. The production method according to claim 1, wherein the aminoacylase of microbial origin is an aminoacylase produced by a strain of the Azotobacter genus.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP8195478A JPS559738A (en) | 1978-07-07 | 1978-07-07 | Production of optically active 3-hydroxy-4- (hydroxymethyl)phenylglycine |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP8195478A JPS559738A (en) | 1978-07-07 | 1978-07-07 | Production of optically active 3-hydroxy-4- (hydroxymethyl)phenylglycine |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS559738A JPS559738A (en) | 1980-01-23 |
| JPS6225359B2 true JPS6225359B2 (en) | 1987-06-02 |
Family
ID=13760885
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP8195478A Granted JPS559738A (en) | 1978-07-07 | 1978-07-07 | Production of optically active 3-hydroxy-4- (hydroxymethyl)phenylglycine |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS559738A (en) |
-
1978
- 1978-07-07 JP JP8195478A patent/JPS559738A/en active Granted
Also Published As
| Publication number | Publication date |
|---|---|
| JPS559738A (en) | 1980-01-23 |
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