JPS5857149B2 - L - Aminosansankakosonoseihou - Google Patents
L - AminosansankakosonoseihouInfo
- Publication number
- JPS5857149B2 JPS5857149B2 JP50153994A JP15399475A JPS5857149B2 JP S5857149 B2 JPS5857149 B2 JP S5857149B2 JP 50153994 A JP50153994 A JP 50153994A JP 15399475 A JP15399475 A JP 15399475A JP S5857149 B2 JPS5857149 B2 JP S5857149B2
- Authority
- JP
- Japan
- Prior art keywords
- sephadex
- activity
- amino acid
- enzyme
- acid oxidase
- 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
- 230000000694 effects Effects 0.000 claims description 24
- 229920005654 Sephadex Polymers 0.000 claims description 19
- 239000012507 Sephadex™ Substances 0.000 claims description 18
- 108010008292 L-Amino Acid Oxidase Proteins 0.000 claims description 10
- 102000007070 L-amino-acid oxidase Human genes 0.000 claims description 10
- 238000000034 method Methods 0.000 claims description 8
- 231100000614 poison Toxicity 0.000 claims description 3
- 239000002574 poison Substances 0.000 claims description 3
- 241000271039 Agkistrodon Species 0.000 claims description 2
- 102000035195 Peptidases Human genes 0.000 claims 1
- 108091005804 Peptidases Proteins 0.000 claims 1
- 235000019833 protease Nutrition 0.000 claims 1
- 102000004190 Enzymes Human genes 0.000 description 14
- 108090000790 Enzymes Proteins 0.000 description 14
- 229940088598 enzyme Drugs 0.000 description 14
- 102000004860 Dipeptidases Human genes 0.000 description 10
- 108090001081 Dipeptidases Proteins 0.000 description 10
- ROHFNLRQFUQHCH-YFKPBYRVSA-N L-leucine Chemical compound CC(C)C[C@H](N)C(O)=O ROHFNLRQFUQHCH-YFKPBYRVSA-N 0.000 description 9
- 239000000872 buffer Substances 0.000 description 8
- 241000142921 Tardigrada Species 0.000 description 7
- 239000002435 venom Substances 0.000 description 7
- 231100000611 venom Toxicity 0.000 description 7
- 210000001048 venom Anatomy 0.000 description 7
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 6
- 229960003136 leucine Drugs 0.000 description 5
- 235000019454 L-leucine Nutrition 0.000 description 4
- 239000004395 L-leucine Substances 0.000 description 4
- 238000000746 purification Methods 0.000 description 4
- 239000007983 Tris buffer Substances 0.000 description 3
- 238000010828 elution Methods 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- 239000003998 snake venom Substances 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- RLFWWDJHLFCNIJ-UHFFFAOYSA-N 4-aminoantipyrine Chemical compound CN1C(C)=C(N)C(=O)N1C1=CC=CC=C1 RLFWWDJHLFCNIJ-UHFFFAOYSA-N 0.000 description 2
- 241001669680 Dormitator maculatus Species 0.000 description 2
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 description 2
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 2
- 102000004316 Oxidoreductases Human genes 0.000 description 2
- 108090000854 Oxidoreductases Proteins 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 229940024606 amino acid Drugs 0.000 description 2
- 238000004040 coloring Methods 0.000 description 2
- 150000002500 ions Chemical class 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- ZIIUUSVHCHPIQD-UHFFFAOYSA-N 2,4,6-trimethyl-N-[3-(trifluoromethyl)phenyl]benzenesulfonamide Chemical compound CC1=CC(C)=CC(C)=C1S(=O)(=O)NC1=CC=CC(C(F)(F)F)=C1 ZIIUUSVHCHPIQD-UHFFFAOYSA-N 0.000 description 1
- BFSVOASYOCHEOV-UHFFFAOYSA-N 2-diethylaminoethanol Chemical compound CCN(CC)CCO BFSVOASYOCHEOV-UHFFFAOYSA-N 0.000 description 1
- 241001464430 Cyanobacterium Species 0.000 description 1
- 102000003886 Glycoproteins Human genes 0.000 description 1
- 108090000288 Glycoproteins Proteins 0.000 description 1
- 108010001336 Horseradish Peroxidase Proteins 0.000 description 1
- FFEARJCKVFRZRR-UHFFFAOYSA-N L-Methionine Natural products CSCCC(N)C(O)=O FFEARJCKVFRZRR-UHFFFAOYSA-N 0.000 description 1
- 150000008575 L-amino acids Chemical class 0.000 description 1
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 1
- 229930195722 L-methionine Natural products 0.000 description 1
- ROHFNLRQFUQHCH-UHFFFAOYSA-N Leucine Natural products CC(C)CC(N)C(O)=O ROHFNLRQFUQHCH-UHFFFAOYSA-N 0.000 description 1
- 102000003992 Peroxidases Human genes 0.000 description 1
- 102000015439 Phospholipases Human genes 0.000 description 1
- 108010064785 Phospholipases Proteins 0.000 description 1
- 239000008351 acetate buffer Substances 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 229940039227 diagnostic agent Drugs 0.000 description 1
- 239000000032 diagnostic agent Substances 0.000 description 1
- 229940079919 digestives enzyme preparation Drugs 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 238000000691 measurement method Methods 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 150000002739 metals Chemical class 0.000 description 1
- 229960004452 methionine Drugs 0.000 description 1
- GUAWMXYQZKVRCW-UHFFFAOYSA-N n,2-dimethylaniline Chemical compound CNC1=CC=CC=C1C GUAWMXYQZKVRCW-UHFFFAOYSA-N 0.000 description 1
- 108040007629 peroxidase activity proteins Proteins 0.000 description 1
- 229960005190 phenylalanine Drugs 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 239000008213 purified water Substances 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 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 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 229960004441 tyrosine Drugs 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Landscapes
- Enzymes And Modification Thereof (AREA)
Description
【発明の詳細な説明】
本発明はカンコクマムシ（Ａｇｋｉ ｓ ｔｒｏｄｏｎ
ｃａｌｉｇｉｎｏｓｕｓ ）毒よりジペプチダーゼ活性
を含！ない純粋なＬ−アミノ酸酸化酵素の精製法にかん
する。DETAILED DESCRIPTION OF THE INVENTION The present invention is directed to the use of Agkis trodon.
caliginosus ) contains dipeptidase activity than the poison! This invention relates to a method for purifying pure L-amino acid oxidase.
更に詳しくはカンコクマムシの顎下線の毒液もしくはこ
のもの＼凍結乾燥標品（蛇毒）をゲルろ週休であるセフ
ァデックスＧ−１００（ファルマシア社製）、イオン交
換体であるＤＥ ＡＥセファデックスＡ−５０（ファル
マシア社製釦よびイオン交換体であるＳＰ−セファデッ
クスＣ−５０（ファルマシア社製）を用いて処理するこ
とを特徴とするジペプチダーゼ活性を含オない純粋なＬ
−アミノ酸酸化酵素を精製する方法にかんする。For more details, gel-filter the venom from the submandibular line of the black tardigrade or this lyophilized sample (snake venom). (Pure L containing no dipeptidase activity characterized by being treated using a button manufactured by Pharmacia and an ion exchanger SP-Sephadex C-50 (manufactured by Pharmacia))
- Concerning a method for purifying amino acid oxidase.
従来より蛇毒中にはプロテアーベホスホリパーゼなどの
酵素が含まれていることが知られてかり、また酵素一般
の精製法の一部として架橋デキストランゲルであるセフ
ァデックスが使用されていることが知られている。It has long been known that snake venom contains enzymes such as proteave phospholipase, and it is also known that Sephadex, a cross-linked dextran gel, is used as part of the purification method for enzymes in general. ing.
しかしながら、カンコクマムシ毒はＬアミノ酸活性が強
力であるので、種々の精製方法が試みられて来たが、未
だに粗酵素標品しか得られて−１らず、シめ）もとの粗
酵素標品中にはジペプチダーゼが含まれていて、血清中
のジペプチダーゼ活性を測定する臨床診断薬として作用
することが不可能であった。However, since C. tardigrade venom has a strong L-amino acid activity, various purification methods have been tried, but so far only crude enzyme preparations have been obtained. The product contained dipeptidase, making it impossible to act as a clinical diagnostic agent for measuring dipeptidase activity in serum.
本発明者らはこのカンコクマムシ毒中の強力なＬ−アミ
ノ酸酸化酵素をジペプチダーゼを含まない純粋な酵素標
品を得る目的で研究を重ねた結果本発明を完成した。The present inventors completed the present invention as a result of repeated research with the aim of obtaining a pure enzyme preparation containing no dipeptidase for the powerful L-amino acid oxidase contained in the venom of the tardigrade.
即ち本発明はカンコクマムシ毒をゲルろ週休であるセフ
ァデックスＧ−１００、イオン交換体であるＤＥＡＥ−
セファデックスおよびイオン交換体であるセファデック
スＣ−５０を用いて処理することよりなるジペプチダー
ゼ活性を含オないＬ−アミノ酸酸化酵素を得る方法であ
る。That is, the present invention uses Sephadex G-100, a gel filter, and DEAE-
This is a method for obtaining an L-amino acid oxidase containing no dipeptidase activity, which comprises treatment with Sephadex and Sephadex C-50, an ion exchanger.
使用される原料としては新鮮なカンコクマムシ毒そのま
＼かもしくは市販の凍結乾燥標品が用いられる。The raw material used is either fresh Canker tardigrade venom or a commercially available freeze-dried preparation.
一般に凍結乾燥標品を緩衝液に溶解して、セファデック
スＧ −１００カラムによる処理を数回くりかえした後
、活性部分を分ホしてＤＥＡＥセファデックスＡ−５０
カラムクロマトグラフイによりＮａｃ、４水のリニアー
グラディエンド溶出を行って活性部分を分増し、更にＳ
Ｐ−セファデックスＣ−５０カラムにて処理してＮａＣ
１水のリニアーグラディエンド溶出によりジペプチダー
ゼ活性を含オない純粋なＬ−アミノ酸酸化酵素を得るこ
とができる。Generally, the lyophilized preparation is dissolved in a buffer solution and treated with a Sephadex G-100 column several times, and then the active portion is separated and treated with a DEAE Sephadex A-50
By column chromatography, linear gradient elution of Nac and 4 water was performed to increase the active portion, and then S
NaC was treated with a P-Sephadex C-50 column.
Pure L-amino acid oxidase containing no dipeptidase activity can be obtained by linear gradient elution with 1 water.
原料の蛇毒の純度によってはセファデックスＧ−１００
やＤＥＡＥ−セファデックスＡ−５０の処理を行わずに
直接ＳＰ−セファデックスＣ−５０カラム処理によって
純粋なＬ−アミノ酸酸化酵素を得ることもできる。Sephadex G-100 depending on the purity of the raw material snake venom.
Pure L-amino acid oxidase can also be obtained by direct treatment with SP-Sephadex C-50 column without treatment with DEAE-Sephadex A-50.
このようにして得られたＬ−アミノ酸酸化酵素の性質を
以下に示す。The properties of the L-amino acid oxidase thus obtained are shown below.
（１）外 観：白色粉末
（２）分子量（ゲルｐ過法） ： ８．５ｘ １０４（
３）等重点：４．９
（４）均一性：ディスク電気泳動的に均一な糖タンパク
質
（５）至適ＰＨ： ８．０−８．５ （第１図参照）Ｐ
Ｈ５，０−７，５で安定。(1) Appearance: White powder (2) Molecular weight (gel p-filtration method): 8.5x 104 (
3) Equal weight: 4.9 (4) Uniformity: disk electrophoretically uniform glycoprotein (5) Optimal pH: 8.0-8.5 (see Figure 1) P
Stable at H5,0-7,5.
（６）熱安定性：５０℃寸で安定 ７０℃、３０分間で失活。(6) Thermal stability: Stable at 50℃ Inactivated at 70°C for 30 minutes.
（７）各種金属による影響 Ｈｇ＋＋：強く阻害される。(7) Effects of various metals Hg++: Strongly inhibited.
Ｆｅ千十 、ｃｏ＋＋９Ｍｎ＋十 、Ｃａ＋＋ 、ｚｎ
＋＋Ｃｕ” ” 、 Ｍｇ” ＋：全く阻害されない。Fe110, co++9Mn+10, Ca++, zn
++Cu"", Mg"+: Not inhibited at all.
（８）基質特異性（第１表参照）
Ｌ−ロイシン、Ｌ−メチオニン、Ｌ−トリプトファン、
Ｌ−フェニルアラニン、Ｌ−チロシンによく作用する。(8) Substrate specificity (see Table 1) L-leucine, L-methionine, L-tryptophan,
It acts well on L-phenylalanine and L-tyrosine.
第１表二種々のアミノ酸に対するＬ−アミノ酸酸化酵素
の活性
ａ）ロイシンに対する活性を１００％とする。Table 1 2 Activity of L-amino acid oxidase towards various amino acids a) Activity towards leucine is taken as 100%.
以下に本発明の実施例を示す。Examples of the present invention are shown below.
実施例 １
１２０ηのカンコクマムシ毒を精製水１０−に溶解し、
０．０５Ｍ、ＮａＣ１を含有する０、０１ＭＴｒｉｓ
ＨＣ１緩衝液（ｐＨ７，６）で緩衝化したセファデック
、’（Ｇ−１００カラム（３，２ｘ ６０Ｃｒｒｌ）に
負荷し、同緩衝液で溶出した。Example 1 Dissolve 120η of tardigrade poison in purified water 10-
0.01M Tris containing 0.05M NaCl
It was loaded onto a Sephadec' (G-100 column (3.2 x 60 Crrl) buffered with HC1 buffer (pH 7.6) and eluted with the same buffer.
この操作で低分子の不純物を除去することができ、酵素
活性の存在する部分（はじめ７５．ら２１６〜２４０−
の流出液）を集めた。This operation makes it possible to remove low-molecular impurities, and also removes enzymatically active parts (Hajime 75. et al. 216-240-
effluent) was collected.
この操作を４回行ない（４８０７ｎＩ！のカンコクマム
シ毒を処理）活性部分を、０．１ ＭＮａＣＡ ｋを
含有する０、０１ Ｍ Ｔｒｉｓ Ｈｃ、ｌ緩衝液（
ｐＨ７，６）で緩衝化したＤＥＡＥ−セファデックスＣ
−５０カラム（１，６５ｘ １５ｃＷＩ）に吸着させ、
同緩衝液でＮａＣｔが０，１Ｍから０．３ Ｍのリニア
ーグラディエンドによシ溶離した。This procedure was repeated four times (4807 nI! of C. tardigrade venom treated) and the active moiety was dissolved in 0.01 M Tris Hc, 1 buffer containing 0.1 M NaCA k.
DEAE-Sephadex C buffered at pH 7.6)
-50 column (1,65x 15cWI),
Elution was performed using the same buffer with a linear gradient of NaCt from 0.1M to 0.3M.
ここで得られた活性部分を濃縮し、０．０４Ｍ酢酸緩衝
液（ｐＨ５，０） ２０−に溶解した。The active portion obtained here was concentrated and dissolved in 0.04M acetate buffer (pH 5,0) 20-.
この酵素液を０．０４Ｍ酢酸緩衝液（ｐＨ５，０）で緩
衝化したｓｐ−セファデックスＣ−５０カラム（１，６
５ｘ１５ｃｒｒｌ）に吸着させ、同緩衝液でＮａＣｔが
ＯＭから０．２のりニア−グラディエンドによう溶離し
た。This enzyme solution was applied to an sp-Sephadex C-50 column (1,6
NaCt was eluted from the OM to a gradient of 0.2 nm using the same buffer.
活性部分を濃縮し、１ＭＴｒｉｓ を加えてｐＨ７，
０に調整し ４℃で保存した。Concentrate the active part and add 1M Tris to pH 7.
0 and stored at 4°C.
以上の精製過程を第■表に示す。The above purification process is shown in Table ①.
活性収率は３８％、カンコクマムン毒から６１倍精製さ
れディスク電気泳動で本酵素は純粋であることが証明さ
れた。The activity yield was 38%, and the enzyme was purified 61 times from Cyanobacterium venom and was proved to be pure by disk electrophoresis.
精製酵素の比活性は３９Ｖ■であう、ジペプチターゼ活
性を示さなかった。The specific activity of the purified enzyme was 39V, and it did not show any dipeptidase activity.
第■表：カンコクマムシ毒よりＬアミノ酸酸化酵素の精
製
精製段階 活性 タンパク 比活性（ｕ）
（７’Ｗ） （ｕＡ’）粗酵素 ３０５４８０ ０
．６３５（１）セファデックス
Ｇ−１ｏｏ１．、．２０３３８．０５．３４（８，４）
１７７デ′り” １４４ ８．３８ １７．２（２
７，１）−５０
ゞ７７デ′り” １１７ ３．００ ３９．０（６
１，４）−５０
実施例 ２
カンコクマムシ毒８６０■を０．０４ Ｍ酢酸緩衝液（
ｐＨ５，０） ２００ｍｌに溶解し、同緩衝液で緩衝化
したＳＰ−セファデックスＣ−５０カラム（１，８Ｘ
２０ＣｒＩＮ）に吸着させ、同緩衝液１ｔでカラムを洗
滌したのち、同緩衝液で、ＮａＣＡがＯＭから０−２
Ｍ ’１でのリニアーグラディエンドで溶離した。Table ■: Purification steps of L-amino acid oxidase from tardigrade venom Activity Protein Specific activity (u)
(7'W) (uA') Crude enzyme 305480 0
．． 635(1) Sephadex G-1oo1. ,．． 20338.05.34 (8,4)
177 days” 144 8.38 17.2 (2
7,1)-50 ゞ77 de'ri" 117 3.00 39.0 (6
1,4)-50 Example 2.
SP-Sephadex C-50 column (1,8X
After washing the column with 1 t of the same buffer, NaCA was reduced from OM to 0-2 CrIN with the same buffer.
It was eluted with a linear gradient at M'1.
第■表に示すように活性収率は５２多であり、比活性１
４．６Ｖ■であり、ジペプチダーゼ活性は認められなか
った。As shown in Table 2, the activity yield was 52%, and the specific activity was 1%.
The voltage was 4.6 V■, and no dipeptidase activity was observed.
（Ｌ−アミノ酸酸化酵素活性測定法）
１憎／−濃度のＬ−ロイシンｌ−に〔４ア□ノアンチピ
リン４７ｑ、Ｎ 、 Ｎ’ ジメチルアニリン１０μを
鮫よび西洋ワサビパーオキシダーゼ４■を５０ＴＩｌの
０．１Ｍ燐酸緩衝液（ｐＨ７，０）に溶解した〕発色液
２ｍｌを加え、酵素０．５−を加え３７℃、１０分反応
後３饅酢酸０．５７ｎｌを加え反応を停止させ５５０
ｎｍにむける吸光度を測定した。(Method for measuring L-amino acid oxidase activity) To L-leucine l- at a concentration of 1%, add 10μ of [4anoantipyrine 47q, N, N' dimethylaniline and 50TIl of horseradish peroxidase 4μ]. Add 2 ml of coloring solution [dissolved in 1M phosphate buffer (pH 7.0)], add 0.5-ml of enzyme and react for 10 minutes at 37°C, then add 0.57 nl of 3-cup acetic acid to stop the reaction.
The absorbance towards nm was measured.
酵素活性は上記反応条件下に１分間に１μｍｏ、／、の
Ｌ−ロイシンを酸化する酵素活性を１単位とした。For the enzyme activity, one unit was defined as the enzyme activity that oxidized 1 μmo/. of L-leucine per minute under the above reaction conditions.
（ジペプチダーゼ活性測定法）
１ｍｔｌｌｒｎｌｌｅｋ度のＬ−ロインルロイシン１−
に、Ｌ−ア□ノ酸酸化酵素（１５ｕ／ｍｇ） ２０ ｐ
Ｌ、〔４−アミノアンチピリン４弘西洋ワサビパーオ
キシダーゼ４ηを５０−の０．０５ Ｍ Ｔｒｉｓ −
ＨＣ，ｇ緩衝液（ｐＨ７，８）に溶解した〕発色液２．
０−に酵素０．１−を加え、３７℃、２０分反応させ、
０．Ｉ Ｎ酢酸１．０−を加え反応を停止させ、５５０
ｎｍの吸光度を測定した。(Dipeptidase activity measurement method) 1mtllrnllek L-loin leleucine 1-
and L-anoic acid oxidase (15u/mg) 20p
L, [4-aminoantipyrine 4-horseradish peroxidase 4η 50-0.05 M Tris-
[Dissolved in HC, g buffer (pH 7, 8)] Coloring solution 2.
Add enzyme 0.1- to 0-, react at 37°C for 20 minutes,
0. The reaction was stopped by adding 1.0-N acetic acid, and 550
The absorbance at nm was measured.
酵素活性は上記反応条件下に２０分間に１μｇのＬ−ロ
イシンを遊離する酵素活性をｌ単位（ｕｎｉｔ）とした
。The enzyme activity was defined as the enzyme activity that releases 1 μg of L-leucine in 20 minutes under the above reaction conditions.
第１図は本酵素のＰＨ安定性を示すグラフ図である。 FIG. 1 is a graph showing the PH stability of this enzyme.
Claims (1)
ｉｇ−ｉｎｏｓｕｓ ）毒をゲルろ週休であるセファデ
ックＧ−１００、イオン交換体であるＤＥＡＥ−セファ
デックスＡ−５０およびイオン交換体であるＳＰ−セフ
ァデックスＣ−５０により処理することを特徴とするジ
ペプチダーゼ活性を含まないＬ−アミノ酸酸化酵素の精
製法。1 Agkistrodon cal
ig-inosus) poison is treated with Sephadex G-100, which is a gel filter, DEAE-Sephadex A-50, which is an ion exchanger, and SP-Sephadex C-50, which is an ion exchanger. A method for purifying L-amino acid oxidase that does not contain peptidase activity.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP50153994A JPS5857149B2 (en) | 1975-12-25 | 1975-12-25 | L - Aminosansankakosonoseihou |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP50153994A JPS5857149B2 (en) | 1975-12-25 | 1975-12-25 | L - Aminosansankakosonoseihou |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS5279079A JPS5279079A (en) | 1977-07-02 |
| JPS5857149B2 true JPS5857149B2 (en) | 1983-12-19 |
Family
ID=15574582
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP50153994A Expired JPS5857149B2 (en) | 1975-12-25 | 1975-12-25 | L - Aminosansankakosonoseihou |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS5857149B2 (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB2122641B (en) * | 1982-06-07 | 1986-08-06 | Otto A Gansow | Metal chelate conjugated monoclonal antibodies where in the metal is an emitter |
-
1975
- 1975-12-25 JP JP50153994A patent/JPS5857149B2/en not_active Expired
Also Published As
| Publication number | Publication date |
|---|---|
| JPS5279079A (en) | 1977-07-02 |
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