JPH0438399B2 - - Google Patents
Info
- Publication number
- JPH0438399B2 JPH0438399B2 JP12926184A JP12926184A JPH0438399B2 JP H0438399 B2 JPH0438399 B2 JP H0438399B2 JP 12926184 A JP12926184 A JP 12926184A JP 12926184 A JP12926184 A JP 12926184A JP H0438399 B2 JPH0438399 B2 JP H0438399B2
- Authority
- JP
- Japan
- Prior art keywords
- amino acid
- carbamyl
- present
- bacterial cells
- yin
- 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
- 150000008575 L-amino acids Chemical class 0.000 claims description 14
- 229940024606 amino acid Drugs 0.000 claims description 14
- 230000001580 bacterial effect Effects 0.000 claims description 11
- 238000004519 manufacturing process Methods 0.000 claims description 9
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 claims description 5
- 244000005700 microbiome Species 0.000 claims description 5
- 150000001413 amino acids Chemical class 0.000 claims description 4
- 229960004441 tyrosine Drugs 0.000 claims description 3
- QDGAVODICPCDMU-UHFFFAOYSA-N 2-amino-3-[3-[bis(2-chloroethyl)amino]phenyl]propanoic acid Chemical compound OC(=O)C(N)CC1=CC=CC(N(CCCl)CCCl)=C1 QDGAVODICPCDMU-UHFFFAOYSA-N 0.000 claims description 2
- IFGCUJZIWBUILZ-UHFFFAOYSA-N sodium 2-[[2-[[hydroxy-(3,4,5-trihydroxy-6-methyloxan-2-yl)oxyphosphoryl]amino]-4-methylpentanoyl]amino]-3-(1H-indol-3-yl)propanoic acid Chemical compound [Na+].C=1NC2=CC=CC=C2C=1CC(C(O)=O)NC(=O)C(CC(C)C)NP(O)(=O)OC1OC(C)C(O)C(O)C1O IFGCUJZIWBUILZ-UHFFFAOYSA-N 0.000 claims description 2
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 claims 1
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 claims 1
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 claims 1
- 229960005190 phenylalanine Drugs 0.000 claims 1
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 claims 1
- 229960004799 tryptophan Drugs 0.000 claims 1
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 claims 1
- 210000004027 cell Anatomy 0.000 description 13
- 238000000034 method Methods 0.000 description 7
- 239000000243 solution Substances 0.000 description 7
- 238000006243 chemical reaction Methods 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 3
- 108090000790 Enzymes Proteins 0.000 description 3
- 229910021380 Manganese Chloride Inorganic materials 0.000 description 3
- GLFNIEUTAYBVOC-UHFFFAOYSA-L Manganese chloride Chemical compound Cl[Mn]Cl GLFNIEUTAYBVOC-UHFFFAOYSA-L 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 238000004166 bioassay Methods 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 3
- 238000004128 high performance liquid chromatography Methods 0.000 description 3
- 235000002867 manganese chloride Nutrition 0.000 description 3
- 239000011565 manganese chloride Substances 0.000 description 3
- 229940099607 manganese chloride Drugs 0.000 description 3
- 235000013372 meat Nutrition 0.000 description 3
- 230000000813 microbial effect Effects 0.000 description 3
- 239000002994 raw material Substances 0.000 description 3
- NWLXJVDJMARXSP-UHFFFAOYSA-N 2-(carbamoylamino)-3-(1h-indol-3-yl)propanoic acid Chemical compound C1=CC=C2C(CC(NC(=O)N)C(O)=O)=CNC2=C1 NWLXJVDJMARXSP-UHFFFAOYSA-N 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 2
- 241000186063 Arthrobacter Species 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- SIKJAQJRHWYJAI-UHFFFAOYSA-N Indole Chemical compound C1=CC=C2NC=CC2=C1 SIKJAQJRHWYJAI-UHFFFAOYSA-N 0.000 description 2
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- -1 N-carbamyl amino Chemical class 0.000 description 2
- 229910002651 NO3 Inorganic materials 0.000 description 2
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 2
- 239000008272 agar Substances 0.000 description 2
- 150000003863 ammonium salts Chemical class 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 239000007795 chemical reaction product Substances 0.000 description 2
- GVPFVAHMJGGAJG-UHFFFAOYSA-L cobalt dichloride Chemical compound [Cl-].[Cl-].[Co+2] GVPFVAHMJGGAJG-UHFFFAOYSA-L 0.000 description 2
- 239000013078 crystal Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 229960004452 methionine Drugs 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- PNLKYZVGQWCHBH-QMMMGPOBSA-N (2s)-2-(carbamoylamino)-3-(4-hydroxyphenyl)propanoic acid Chemical compound NC(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 PNLKYZVGQWCHBH-QMMMGPOBSA-N 0.000 description 1
- DEWDMTSMCKXBNP-UHFFFAOYSA-N 2-(carbamoylamino)-4-methylsulfanylbutanoic acid Chemical compound CSCCC(C(O)=O)NC(N)=O DEWDMTSMCKXBNP-UHFFFAOYSA-N 0.000 description 1
- 241000186073 Arthrobacter sp. Species 0.000 description 1
- 102000016938 Catalase Human genes 0.000 description 1
- 108010053835 Catalase Proteins 0.000 description 1
- RWSOTUBLDIXVET-UHFFFAOYSA-N Dihydrogen sulfide Chemical compound S RWSOTUBLDIXVET-UHFFFAOYSA-N 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 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 description 1
- 238000003794 Gram staining Methods 0.000 description 1
- FFEARJCKVFRZRR-UHFFFAOYSA-N L-Methionine Natural products CSCCC(N)C(O)=O FFEARJCKVFRZRR-UHFFFAOYSA-N 0.000 description 1
- 229930195722 L-methionine Natural products 0.000 description 1
- 240000006024 Lactobacillus plantarum Species 0.000 description 1
- 235000013965 Lactobacillus plantarum Nutrition 0.000 description 1
- 238000005481 NMR spectroscopy Methods 0.000 description 1
- 102000004316 Oxidoreductases Human genes 0.000 description 1
- 108090000854 Oxidoreductases Proteins 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 108010046334 Urease Proteins 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 125000003917 carbamoyl group Chemical group [H]N([H])C(*)=O 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 210000003495 flagella Anatomy 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 229940091173 hydantoin Drugs 0.000 description 1
- 150000001469 hydantoins Chemical class 0.000 description 1
- 229910000037 hydrogen sulfide Inorganic materials 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 230000003301 hydrolyzing effect Effects 0.000 description 1
- PZOUSPYUWWUPPK-UHFFFAOYSA-N indole Natural products CC1=CC=CC2=C1C=CN2 PZOUSPYUWWUPPK-UHFFFAOYSA-N 0.000 description 1
- RKJUIXBNRJVNHR-UHFFFAOYSA-N indolenine Natural products C1=CC=C2CC=NC2=C1 RKJUIXBNRJVNHR-UHFFFAOYSA-N 0.000 description 1
- 238000009630 liquid culture Methods 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 235000019341 magnesium sulphate Nutrition 0.000 description 1
- 229930182817 methionine Natural products 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 235000019796 monopotassium phosphate Nutrition 0.000 description 1
- 230000004899 motility Effects 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- NNFCIKHAZHQZJG-UHFFFAOYSA-N potassium cyanide Chemical compound [K+].N#[C-] NNFCIKHAZHQZJG-UHFFFAOYSA-N 0.000 description 1
- 238000004062 sedimentation Methods 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- UQDJGEHQDNVPGU-UHFFFAOYSA-N serine phosphoethanolamine Chemical compound [NH3+]CCOP([O-])(=O)OCC([NH3+])C([O-])=O UQDJGEHQDNVPGU-UHFFFAOYSA-N 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 235000000346 sugar Nutrition 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 238000004809 thin layer chromatography Methods 0.000 description 1
- 238000002604 ultrasonography Methods 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Description
〔産業上の利用分野〕
本発明は、アミノ酸のN−カルバミル体から微
生物酵素系を利用して相当するL−アミノ酸を製
造する方法に関する。
〔従来の技術〕
従来、微生物酵素系を利用したDL−N−カル
バミルアミノ酸よりのL−アミノ酸の製造方法に
ついては、DL−N−カルバミルメチオニンより
のL−メチオニンよりの製造法が知られている
(特公昭55−29678号)。
〔発明が解決しようとする問題点〕
従来の方法では、D−N−カルバミルメチオニ
ンはそのまま残存するため、反応生成液からのL
−メチオニンとD−N−カルバミルメチオニンと
の分離を必要とする。
本発明の目的は、D体及びL体に関係なくすべ
てのN−カルバミルアミノ酸を、微生物酵素系を
利用して相当するL−アミノ酸に変換する、L−
アミノ酸の製造方法を提供することにある。
〔問題点を解決するための手段〕
本発明を概説すれば、本発明はL−アミノ酸の
製造方法に関する発明であつて、フエニルアラニ
ン、トリプトフアン、及びチロシンよりなる群か
ら選択したアミノ酸のN−カルバミル体のL体、
D体又はDL体に、アリスロバクター属微生物の
培養液、菌体又は菌体処理物を作用させ、生成し
た相当するL−フエニルアラニン、L−トリプト
フアン、又はL−チロシンを採取することを特徴
とする。
本発明方法によれば、D型のN−カルバミル体
も、相当するL−アミノ酸に変換することができ
る。
本発明方法で使用する原料の例としては、N−
カルバミルフエニルアラニン、N−カルバミルト
リプトフアン及びN−カルバミルチロシン等が挙
げられる。これらは、D、L及びDL体のいずれ
のものであつてよい。
本発明方法の原料は、公知の物質であるか、公
知の方法によつて得ることができるものであり、
例えばDL−N−カルバミルアミノ酸は、DL−ア
ミノ酸にシアン化カリウム等を反応させる公知の
方法で、またアミノ酸のDL−ヒダントイン誘導
体を適当な条件で加水分解する方法で得ることが
できる。
次に本発明方法で使用するアリスロバクター属
(Arthrobacter)微生物としては、その培養液を
そのまま用いることもできるが、生菌体、凍結菌
体、凍結乾燥菌体又は菌体磨砕物若しくは菌体抽
出物のような菌体処理物を使用してもよい。ま
た、変異処理により性能の向上した変異株など
も、より効果的に使用できることはいうまでもな
い。
上記アリスロバクター属微生物のうちで好適な
例としては、本発明者等が自然界から新たに分離
した新菌種であるアリスロバクター属DK−200
菌株がある。なお、該DK−200菌株は、
Arthrobacter sp.DK−200(アリスロバクター属)
と表示して、工業技術院微生物工業技術研究所
に、微工研菌寄第7472号(FERM P−7472)と
して寄託されている。
本発明者等は、先に上記アリスロバクター属
DK−200菌株の培養物等を用いて、5−置換ヒ
ダントインから相当するL−アミノ酸を製造する
方法を開発した(特願昭59−70906号)。
以下、上記DK−200菌株の菌学的諸性質を示
す。
1 形態
(1) 細胞の形及び大きさ: 0.3〜0.5×0.8〜5.0μm
(2) 多形性の有無: 有
(3) 運動性の有無: 無
(4) 鞭毛の有無、着生状態 無
(5) 胞子の有無: 無
(6) グラム染色性:
陰性〜弱陽性であるがグラム陽性粒子を有する
(7) 抗酸性: 無
2 各培地での生育状態
(1) 肉汁寒天平板培養
コロニーの色: 淡白色
コロニーの形状: 円形
コロニーの隆起: 凸
コロニーの周縁: 全縁
(2) 肉汁寒天斜面培養
生育状態: 適度
光 沢: 有
形 状: 糸状
(3) 肉汁液体培養
混濁の程度: 均一
沈 殿: 有
液面での生育: 特に無
(4) 肉汁ゼラチン穿刺培養: 液化する
3 生理学的性質
(1) 硝酸塩の還元: 陰
(2) 脱窒反応: 陽
(3) MRテスト: 陰
(4) VPテスト: 陰
(5) インドールの生成: 陰
(6) 硫化水素の生成: 陰
(7) デンプンの加水分解: 陰
(8) クエン酸の利用:
コーサー(Koser)培地及びクリステンセン
(Christensen)培地共利用しない
(9) 無機窒素源の利用: 硝酸塩及びアンモニウム
塩共利用する
(10) 色素の生成: 無
(11) ウレアーゼ: 陰
(12) オキシダーゼ: 陰
(13) カタラーゼ: 陽
(14) 酸素に対する態度: 好気性
(15) 生育の範囲
温度: 17.8〜33.2℃
PH: 5〜10
(16) OFテスト: 酸化的(oxidative)
(17) 糖類からの酸及びガスの生成の有無
[Industrial Application Field] The present invention relates to a method for producing a corresponding L-amino acid from an N-carbamyl form of an amino acid using a microbial enzyme system. [Prior Art] Conventionally, as a method for producing L-amino acid from DL-N-carbamyl amino acid using a microbial enzyme system, a method for producing L-methionine from DL-N-carbamylmethionine is known. (Special Publication No. 55-29678). [Problems to be solved by the invention] In the conventional method, since DN-carbamylmethionine remains as it is, L from the reaction product solution is
- Requires separation of methionine and DN-carbamylmethionine. The purpose of the present invention is to convert all N-carbamyl amino acids, regardless of D and L forms, into the corresponding L-amino acids using microbial enzyme systems.
An object of the present invention is to provide a method for producing amino acids. [Means for Solving the Problems] To summarize the present invention, the present invention relates to a method for producing an L-amino acid, and the present invention relates to a method for producing an L-amino acid. L-form of carbamyl body,
The D-form or DL-form is treated with a culture solution, bacterial cells, or treated bacterial cells of a microorganism belonging to the genus Arilobacter, and the corresponding L-phenylalanine, L-tryptophan, or L-tyrosine produced is collected. Features. According to the method of the present invention, the D-type N-carbamyl form can also be converted into the corresponding L-amino acid. Examples of raw materials used in the method of the present invention include N-
Examples include carbamylphenylalanine, N-carbamyltryptophan, and N-carbamyltyrosine. These may be any of the D, L and DL forms. The raw material for the method of the present invention is a known substance or can be obtained by a known method,
For example, DL-N-carbamyl amino acid can be obtained by a known method of reacting DL-amino acid with potassium cyanide or the like, or by hydrolyzing a DL-hydantoin derivative of an amino acid under appropriate conditions. Next, as the Arthrobacter microorganism used in the method of the present invention, its culture solution can be used as it is, but live cells, frozen cells, freeze-dried cells, ground bacterial cells, or bacterial cells can be used. A processed product such as an extract may also be used. It goes without saying that mutant strains with improved performance through mutation treatment can also be used more effectively. Among the microorganisms of the genus Arithrobacter mentioned above, a suitable example is Arilobacter genus DK-200, which is a new bacterial species newly isolated from the natural world by the present inventors.
There are strains. In addition, the DK-200 strain is
Arthrobacter sp.DK−200 (Arthrobacter genus)
It has been deposited with the National Institute of Microbiology, Agency of Industrial Science and Technology as FERM P-7472. The present inventors previously discovered that the above-mentioned Arilobacter spp.
A method for producing the corresponding L-amino acid from 5-substituted hydantoin was developed using a culture of the DK-200 strain (Japanese Patent Application No. 70906/1983). The mycological properties of the DK-200 strain described above are shown below. 1 Morphology (1) Cell shape and size: 0.3 to 0.5 x 0.8 to 5.0 μm (2) Presence or absence of pleomorphism: Yes (3) Presence or absence of motility: No (4) Presence or absence of flagella, epiphytic state No (5) Presence of spores: None (6) Gram staining:
Negative to weakly positive, but has Gram-positive particles (7) Acid-fast: None Growth status on each medium (1) Broth agar plate culture Colony color: Pale white Colony shape: Round Colony bulge: Convex Colony Periphery: Entire edge (2) Meat juice agar slant culture Growth condition: Moderate Gloss: Yes Shape: Thread-like (3) Meat juice liquid culture Degree of turbidity: Uniform Sedimentation: Yes Growth on liquid surface: None (4) Meat juice gelatin puncture culture: Liquefaction 3 Physiological properties (1) Nitrate reduction: Yin (2) Denitrification reaction: Yang (3) MR test: Yin (4) VP test: Yin (5) Indole formation: Yin ( 6) Formation of hydrogen sulfide: Yin (7) Hydrolysis of starch: Yin (8) Utilization of citric acid:
Do not use Koser medium or Christensen medium (9) Use of inorganic nitrogen source: Use nitrate and ammonium salts (10) Pigment production: None (11) Urease: Yin (12) Oxidase: Yin (13) Catalase: Positive (14) Attitude towards oxygen: Aerobic (15) Growth range Temperature: 17.8-33.2°C PH: 5-10 (16) OF test: Oxidative (17) Acid from sugars and presence or absence of gas generation.
【表】【table】
以下、本発明を実施例により更に具体的に説明
するが、本発明はこれらに限定されるものではな
い。なお、特に断わらない限り、各例中の%は重
量%である。
実施例 1
グルコース0.5%、酵母エキス0.2%、リン酸水
素二ナトリウム0.1%、リン酸二水素カリウム
0.05%、硫酸マグネシウム0.05%を含む培地(PH
7.0)5mlを試験管に分注し、120℃で15分間滅菌
した。これに別に殺菌したDL−N−カルバミル
トリプトフアンを、最終濃度が0.1%になるよう
に添加した後、アリスロバクター属DK−200菌
を接種し、30℃で24時間振とう培養した。この培
養液より菌体を遠心分離して集め、同量の生理食
塩水で洗浄後、各別に下記第1表に記載のL−N
−カルバミルアミノ酸0.1M及び塩化マンガン
0.1mMを含む0.1Mトリス塩酸緩衝液(PH7.5)5
ml中に懸濁させ、35℃で20時間反応させた。目的
生成物をバイオアツセイ〔ラクトバチルス・アラ
ビノザス(Lactobacillus arabinosus)ATCC
8014を用いた〕とHPLCにより定量した結果を第
1表に示す。両分析値が一致したことから、いず
れの場合も、目的生成物は相当するL−アミノ酸
と確認した。
EXAMPLES Hereinafter, the present invention will be explained in more detail with reference to Examples, but the present invention is not limited thereto. In addition, unless otherwise specified, % in each example is weight %. Example 1 Glucose 0.5%, yeast extract 0.2%, disodium hydrogen phosphate 0.1%, potassium dihydrogen phosphate
0.05%, magnesium sulfate 0.05% (PH
7.0) 5 ml was dispensed into test tubes and sterilized at 120°C for 15 minutes. After separately sterilized DL-N-carbamyltryptophan was added to the final concentration of 0.1%, Arilobacter DK-200 was inoculated and cultured with shaking at 30°C for 24 hours. The bacterial cells were collected by centrifugation from this culture solution, washed with the same amount of physiological saline, and then separated into L-N cells listed in Table 1 below.
- Carbamyl amino acid 0.1M and manganese chloride
0.1M Tris-HCl buffer (PH7.5) containing 0.1mM
ml and reacted at 35°C for 20 hours. Bioassay the desired product [Lactobacillus arabinosus ATCC]
Table 1 shows the results of quantitative determination by HPLC. Since both analytical values matched, the target product was confirmed to be the corresponding L-amino acid in both cases.
【表】
実施例 2
培地100mlを含む500ml容の三角フラスコを用い
る以外は実施例1と同様に培養して得た菌体を、
下記第2表に記載のD−N−カルバミルアミノ酸
0.1M、塩化コバルト0.5mM、及び塩化マンガン
1mMを含む0.1Mトリス塩酸緩衝液(PH7.0)100
ml中に懸濁させ、35℃で60時間反応させた。反応
生成液のHPLC分析によれば、反応はほぼ完結し
ていた。遠心分離により除菌後の上清を25mlまで
濃縮し、エタノール約5mlを加え、冷却(5℃)
して結晶を得た。これを再結晶し、乾燥して、
HPLC、TLC、バイオアツセイ、NMR及び旋光
度分析を行つたところ、第2表に示すように、各
相当するL−アミノ酸の標品と一致することを確
認した。[Table] Example 2 Bacterial cells obtained by culturing in the same manner as in Example 1 except that a 500 ml Erlenmeyer flask containing 100 ml of medium was used.
D-N-carbamyl amino acids listed in Table 2 below
0.1M, cobalt chloride 0.5mM, and manganese chloride
0.1M Tris-HCl buffer (PH7.0) containing 1mM 100
ml and reacted at 35°C for 60 hours. According to HPLC analysis of the reaction product liquid, the reaction was almost complete. Concentrate the supernatant after sterilization to 25 ml by centrifugation, add approximately 5 ml of ethanol, and cool (5°C).
and obtained crystals. This is recrystallized and dried,
HPLC, TLC, bioassay, NMR, and optical rotation analysis were performed, and as shown in Table 2, it was confirmed that each sample corresponded to the corresponding L-amino acid standard.
【表】
実施例 3
実施例2におけるD−N−カルバミルアミノ酸
をDL−N−カルバミルアミノ酸に変えた以外は
実施例2と同様な操作を行つて、相当するL−ア
ミノ酸の結晶を得た。これらの分析結果は、標品
のL−アミノ酸と一致した。結果を下記第表3に
示す。[Table] Example 3 The same procedure as in Example 2 was performed except that DN-carbamyl amino acid in Example 2 was changed to DL-N-carbamyl amino acid to obtain crystals of the corresponding L-amino acid. Ta. These analysis results were consistent with the standard L-amino acid. The results are shown in Table 3 below.
【表】
実施例 4
実施例2と同様にして得た洗浄菌体を、塩化コ
バルト0.5mMと塩化マンガン1mMとを含む0.1M
トリス塩酸緩衝液(PH7.0)20ml中に懸濁させた。
この懸濁液を、氷冷下において、20kHzの超音波
で3分間3回処理して、菌体破砕液を調製した。
この液に、DL−N−カルバミルアミノ酸を0.1M
になるように添加し、30℃で20時間反応させた。
反応後、不溶物を遠心除去して得た上清を、
HLPC及びバイオアツセイした結果、生成物は、
相当するL−アミノ酸と確認した。分析結果を、
下記第4表に示す。[Table] Example 4 Washed bacterial cells obtained in the same manner as in Example 2 were mixed with 0.1M solution containing 0.5mM cobalt chloride and 1mM manganese chloride.
It was suspended in 20 ml of Tris-HCl buffer (PH7.0).
This suspension was treated with 20 kHz ultrasound three times for 3 minutes under ice cooling to prepare a bacterial cell disruption solution.
Add 0.1M DL-N-carbamyl amino acid to this solution.
The mixture was added so as to give a reaction temperature of 30.degree. C. and reacted at 30.degree. C. for 20 hours.
After the reaction, the supernatant obtained by centrifuging to remove insoluble matter,
As a result of HLPC and bioassay, the product is
It was confirmed to be the corresponding L-amino acid. The analysis results,
It is shown in Table 4 below.
【表】
〔発明の効果〕
以上説明したように、本発明によれば、N−カ
ルバミルアミノ酸から、アリスロバクター属微生
物を使用して、原料がD−、L−又はDL−体で
あることに関係なく、効率良く、相当するL−ア
ミノ酸を製造することができるという顕著な効果
が奏せられる。[Table] [Effects of the Invention] As explained above, according to the present invention, the raw material is D-, L- or DL-form from N-carbamyl amino acid using microorganisms of the genus Arilobacter. Regardless, the remarkable effect of being able to efficiently produce the corresponding L-amino acid can be achieved.
Claims (1)
ロシンよりなる群から選択したアミノ酸のN−カ
ルバミル体のD体、L体又はDL体に、アリスロ
バクター属微生物の培養液、菌体又は菌体処理物
を作用させ、生成した相当するL−フエニルアラ
ニン、L−トリプトフアン、又はL−チロシンを
採取することを特徴とするL−アミノ酸の製造方
法。1. A culture solution, bacterial cells, or treated bacterial cells of a microorganism belonging to the genus Arilobacter is applied to the D, L, or DL form of the N-carbamyl form of an amino acid selected from the group consisting of phenylalanine, tryptophan, and tyrosine. 1. A method for producing an L-amino acid, which comprises collecting the corresponding L-phenylalanine, L-tryptophan, or L-tyrosine produced.
Priority Applications (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP12926184A JPS619293A (en) | 1984-06-25 | 1984-06-25 | Production of l-amino acid |
DK161085A DK164923C (en) | 1984-04-11 | 1985-04-10 | METHOD OF PREPARING L-ALFA AMINO ACIDS |
DE8585302548T DE3582354D1 (en) | 1984-04-11 | 1985-04-11 | METHOD FOR PRODUCING L-AMINO ACIDS. |
EP85302548A EP0159866B1 (en) | 1984-04-11 | 1985-04-11 | Process for production of l-amino acids |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP12926184A JPS619293A (en) | 1984-06-25 | 1984-06-25 | Production of l-amino acid |
Publications (2)
Publication Number | Publication Date |
---|---|
JPS619293A JPS619293A (en) | 1986-01-16 |
JPH0438399B2 true JPH0438399B2 (en) | 1992-06-24 |
Family
ID=15005191
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP12926184A Granted JPS619293A (en) | 1984-04-11 | 1984-06-25 | Production of l-amino acid |
Country Status (1)
Country | Link |
---|---|
JP (1) | JPS619293A (en) |
Families Citing this family (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6780443B1 (en) | 2000-02-04 | 2004-08-24 | Takasago International Corporation | Sensate composition imparting initial sensation upon contact |
US20190054200A1 (en) | 2016-02-24 | 2019-02-21 | Takasago International Corporation | Household product delivering warming and/or tingling sensations |
-
1984
- 1984-06-25 JP JP12926184A patent/JPS619293A/en active Granted
Also Published As
Publication number | Publication date |
---|---|
JPS619293A (en) | 1986-01-16 |
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