JPS5915639B2 - Method for measuring ornithine or ornithine aminotransferase - Google Patents
Method for measuring ornithine or ornithine aminotransferaseInfo
- Publication number
- JPS5915639B2 JPS5915639B2 JP421377A JP421377A JPS5915639B2 JP S5915639 B2 JPS5915639 B2 JP S5915639B2 JP 421377 A JP421377 A JP 421377A JP 421377 A JP421377 A JP 421377A JP S5915639 B2 JPS5915639 B2 JP S5915639B2
- Authority
- JP
- Japan
- Prior art keywords
- ornithine
- measuring
- pyrroline
- aminotransferase
- carboxylic acid
- 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
- AHLPHDHHMVZTML-BYPYZUCNSA-N L-Ornithine Chemical compound NCCC[C@H](N)C(O)=O AHLPHDHHMVZTML-BYPYZUCNSA-N 0.000 title claims description 12
- AHLPHDHHMVZTML-UHFFFAOYSA-N Orn-delta-NH2 Natural products NCCCC(N)C(O)=O AHLPHDHHMVZTML-UHFFFAOYSA-N 0.000 title claims description 11
- 229960003104 ornithine Drugs 0.000 title claims description 11
- UTJLXEIPEHZYQJ-UHFFFAOYSA-N Ornithine Natural products OC(=O)C(C)CCCN UTJLXEIPEHZYQJ-UHFFFAOYSA-N 0.000 title claims description 10
- 238000000034 method Methods 0.000 title claims description 10
- 102000004132 Ornithine aminotransferases Human genes 0.000 title claims description 7
- 108090000691 Ornithine aminotransferases Proteins 0.000 title claims description 7
- 239000005515 coenzyme Substances 0.000 claims description 13
- 150000003222 pyridines Chemical class 0.000 claims description 11
- NXOIMAMHRHDCFR-UHFFFAOYSA-N 2,3-dihydro-1h-pyrrole-2-carboxylic acid Chemical compound OC(=O)C1CC=CN1 NXOIMAMHRHDCFR-UHFFFAOYSA-N 0.000 claims description 10
- 238000012360 testing method Methods 0.000 claims description 7
- KPGXRSRHYNQIFN-UHFFFAOYSA-N 2-oxoglutaric acid Chemical compound OC(=O)CCC(=O)C(O)=O KPGXRSRHYNQIFN-UHFFFAOYSA-N 0.000 claims description 4
- 238000002835 absorbance Methods 0.000 claims description 3
- BAWFJGJZGIEFAR-NNYOXOHSSA-N NAD zwitterion Chemical group NC(=O)C1=CC=C[N+]([C@H]2[C@@H]([C@H](O)[C@@H](COP([O-])(=O)OP(O)(=O)OC[C@@H]3[C@H]([C@@H](O)[C@@H](O3)N3C4=NC=NC(N)=C4N=C3)O)O2)O)=C1 BAWFJGJZGIEFAR-NNYOXOHSSA-N 0.000 claims description 2
- 229930027945 nicotinamide-adenine dinucleotide Natural products 0.000 claims description 2
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 claims 2
- XJLXINKUBYWONI-NNYOXOHSSA-N NADP zwitterion Chemical compound NC(=O)C1=CC=C[N+]([C@H]2[C@@H]([C@H](O)[C@@H](COP([O-])(=O)OP(O)(=O)OC[C@@H]3[C@H]([C@@H](OP(O)(O)=O)[C@@H](O3)N3C4=NC=NC(N)=C4N=C3)O)O2)O)=C1 XJLXINKUBYWONI-NNYOXOHSSA-N 0.000 claims 1
- 229950006238 nadide Drugs 0.000 claims 1
- UMJSCPRVCHMLSP-UHFFFAOYSA-N pyridine Natural products COC1=CC=CN=C1 UMJSCPRVCHMLSP-UHFFFAOYSA-N 0.000 claims 1
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 12
- 239000008057 potassium phosphate buffer Substances 0.000 description 9
- 230000000694 effects Effects 0.000 description 8
- 102000004190 Enzymes Human genes 0.000 description 6
- 108090000790 Enzymes Proteins 0.000 description 6
- 239000000243 solution Substances 0.000 description 6
- 239000006228 supernatant Substances 0.000 description 6
- 238000006243 chemical reaction Methods 0.000 description 5
- 239000000843 powder Substances 0.000 description 5
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 description 4
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 3
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 3
- 238000010438 heat treatment Methods 0.000 description 3
- 210000004185 liver Anatomy 0.000 description 3
- 238000005259 measurement Methods 0.000 description 3
- 238000003756 stirring Methods 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- GUBGYTABKSRVRQ-WFVLMXAXSA-N DEAE-cellulose Chemical compound OC1C(O)C(O)C(CO)O[C@H]1O[C@@H]1C(CO)OC(O)C(O)C1O GUBGYTABKSRVRQ-WFVLMXAXSA-N 0.000 description 2
- 239000000287 crude extract Substances 0.000 description 2
- 239000008187 granular material Substances 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 239000004570 mortar (masonry) Substances 0.000 description 2
- 239000001103 potassium chloride Substances 0.000 description 2
- 235000011164 potassium chloride Nutrition 0.000 description 2
- 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 1
- VDVJQFLGCFFFBP-UHFFFAOYSA-N 2,3-dihydropyrrole-1-carboxylic acid Chemical compound OC(=O)N1CCC=C1 VDVJQFLGCFFFBP-UHFFFAOYSA-N 0.000 description 1
- RSEBUVRVKCANEP-UHFFFAOYSA-N 2-pyrroline Chemical compound C1CC=CN1 RSEBUVRVKCANEP-UHFFFAOYSA-N 0.000 description 1
- XJLXINKUBYWONI-NNYOXOHSSA-O NADP(+) Chemical compound NC(=O)C1=CC=C[N+]([C@H]2[C@@H]([C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OC[C@@H]3[C@H]([C@@H](OP(O)(O)=O)[C@@H](O3)N3C4=NC=NC(N)=C4N=C3)O)O2)O)=C1 XJLXINKUBYWONI-NNYOXOHSSA-O 0.000 description 1
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 description 1
- 239000008351 acetate buffer Substances 0.000 description 1
- HWXBTNAVRSUOJR-UHFFFAOYSA-N alpha-hydroxyglutaric acid Natural products OC(=O)C(O)CCC(O)=O HWXBTNAVRSUOJR-UHFFFAOYSA-N 0.000 description 1
- 229940009533 alpha-ketoglutaric acid Drugs 0.000 description 1
- 230000008033 biological extinction Effects 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 239000000356 contaminant Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 239000003456 ion exchange resin Substances 0.000 description 1
- 229920003303 ion-exchange polymer Polymers 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 229940101270 nicotinamide adenine dinucleotide (nad) Drugs 0.000 description 1
- BOPGDPNILDQYTO-NNYOXOHSSA-N nicotinamide-adenine dinucleotide Chemical compound C1=CCC(C(=O)N)=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](COP(O)(=O)OP(O)(=O)OC[C@@H]2[C@H]([C@@H](O)[C@@H](O2)N2C3=NC=NC(N)=C3N=C2)O)O1 BOPGDPNILDQYTO-NNYOXOHSSA-N 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000012264 purified product Substances 0.000 description 1
- ZVJHJDDKYZXRJI-UHFFFAOYSA-N pyrroline Natural products C1CC=NC1 ZVJHJDDKYZXRJI-UHFFFAOYSA-N 0.000 description 1
- 238000011160 research Methods 0.000 description 1
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Description
【発明の詳細な説明】
本発明はオルニチン・アミノ基転移酵素（以下ＯＴＡと
称する）またはオルニチンの測定法に関する。DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method for measuring ornithine aminotransferase (hereinafter referred to as OTA) or ornithine.
２０従来、ＯＴＡの活性を測定する方法としては、次の
反応式ピロリンー５−カルボン酸
−ノ
ーヘ
グルタメート
るが、反応速度が遅く、終濃度１０゜ｍＭの如き高濃度
の０−アミノベンツアルデヒドを必要とする、モル吸光
係数が２．７１×１０３と低いなどの欠点を有する。20 The conventional method for measuring the activity of OTA is the following reaction formula: pyrroline-5-carboxylic acid-noheglutamate, but the reaction rate is slow and requires a high concentration of 0-aminobenzaldehyde, such as a final concentration of 10 mm. It has drawbacks such as a low molar extinction coefficient of 2.71×10 3 .
そこで本発明者は斯る欠点を除去せんと種々研３５究を
行つた結果、上記ピロリンー５−カルボン酸を還元酵素
でプロリンに変化させ、その際のピリジンヌクレオチド
補酵素の酸化を測定することによつて０ＴＡ活性または
オルニチンを測定する方法を見出した。Therefore, the present inventor conducted various research in an attempt to eliminate such drawbacks, and as a result, decided to convert the above pyrroline-5-carboxylic acid into proline using a reductase and measure the oxidation of the pyridine nucleotide coenzyme at that time. Therefore, we have found a method for measuring OTA activity or ornithine.
更にまた、本発明者は、この測定のために使用される有
利な性質をもつたピロリン−カルボン酸還元酵素がＳａ
ｃｃｈａｒＯｍｙｃｅｓ中に存在することを見出し、こ
れを比較的簡単に精製する方法を見出した。従つて、本
発明は、１α−ケトグルタル酸、還元型ピリジンヌクレ
オチド補酵素、オルニチンおよびピロリン−５−カルボ
ン酸還元酵素からなる系に被検試料を加えて反応させ、
該ピリジンヌク薔レオチド補酵素の減少を測定して０Ｔ
Ａ活性を測定する方法と２α−ケトグルタル酸、還元型
ピリジンヌクレオチド補酵素、０ＴＡおよびピロリン＝
５−カルボン酸還元酵素からなる系に被検試料を加えて
反応させ、該ピリジンヌクレオチド補酵素の減少を測定
してオルニチンを測定する方法である。Furthermore, the inventors have discovered that the pyrroline-carboxylic acid reductase with advantageous properties used for this measurement is Sa
ccharOmyces, and found a relatively simple method to purify it. Therefore, the present invention involves adding a test sample to a system consisting of 1α-ketoglutarate, reduced pyridine nucleotide coenzyme, ornithine, and pyrroline-5-carboxylic acid reductase, and causing a reaction.
0T by measuring the decrease in the pyridine-reotide coenzyme.
Method for measuring A activity and 2α-ketoglutarate, reduced pyridine nucleotide coenzyme, OTA and pyrroline =
In this method, a test sample is added to a system consisting of 5-carboxylic acid reductase and reacted, and ornithine is measured by measuring the decrease in the pyridine nucleotide coenzyme.
本発明の反応機構を式で示せぱ次のとうりである。The reaction mechanism of the present invention can be expressed as follows.
本発明におけるピリジンヌクレオチド補酵素としては、
例えばニコチンアミドアデニンジヌクレオチド（ＮＡＤ
）またはニコチンアミドアデニンジヌクレオチドリン酸
（ＮＡＤＰ）が使用される。The pyridine nucleotide coenzyme in the present invention includes:
For example, nicotinamide adenine dinucleotide (NAD
) or nicotinamide adenine dinucleotide phosphate (NADP) is used.
ピロリン−５−カルボン酸還元酵素としては今回本発明
者によつてパン酵母（ＳａｃｃｈａｒＯｍｙｃｅｓｃｅ
ｒｅｖｉｓｉａｅ）から濃縮精製されたものを使用する
のが好ましい。当該酵素は後述の実施例１に記載の方法
によつて、パン酵母をアセトン処理してアセトン粉末と
なし、これを水に懸濁して酵素を抽出し、次いでイオン
交換樹脂処理、熱処理を行つて約１５０倍に濃縮精製す
ることができる。この還元酵素は０ＴＡの数倍ないし１
０倍量使用することが必要である。本発明方法を実施す
るには、α−ケトグルタル酸、還元型ピリジンヌクレオ
チド補酵素、ピロリン−５−カルボン酸還元酵素および
オルニチンまたは０ＴＡの何れか一つを含む系にオルニ
チンまたは０ＴＡを含む被検試料を加えて反応させ、還
元型ピリジンヌクレオチド補酵素の減少を測定する。As the pyrroline-5-carboxylic acid reductase, the present inventor has developed baker's yeast (SaccharOmycesce).
It is preferable to use concentrated and purified products from A. revisiae). The enzyme was obtained by treating baker's yeast with acetone to obtain acetone powder, suspending it in water to extract the enzyme, and then subjecting it to ion exchange resin treatment and heat treatment. It can be concentrated and purified approximately 150 times. This reductase is several times that of 0TA to 1
It is necessary to use 0 times the amount. To carry out the method of the present invention, a test sample containing ornithine or OTA in a system containing α-ketoglutarate, reduced pyridine nucleotide coenzyme, pyrroline-5-carboxylic acid reductase, and either ornithine or OTA is used. is added to react, and the decrease in reduced pyridine nucleotide coenzyme is measured.
当該補酵素としてＮＡＤＨを使用したときは約３７℃の
温度で反応させ、３４０ｎｍにおける吸光度の減少を測
定することによりオルニチンまたは０ＴＡを測定するこ
とができる。When NADH is used as the coenzyme, ornithine or 0TA can be measured by reacting at a temperature of about 37° C. and measuring the decrease in absorbance at 340 nm.
実施例 １
ピロリン−５−カルボン酸還元酵素の調製：一２０℃に
凍結保存しておいたパン酵母５００９を４℃の冷蔵庫中
に一夜放置して融解し、濾紙上に薄く広げて室温で６時
間風乾し乾燥させる。Example 1 Preparation of pyrroline-5-carboxylic acid reductase: Baker's yeast 5009, which had been frozen and stored at -20°C, was left in a refrigerator at 4°C overnight to thaw, spread thinly on a filter paper, and incubated at room temperature for 60 minutes. Air dry for an hour.
乳鉢で微細顆粒状になるまで粉砕し、乾燥粉末約１７０
ｙを得る。これに−２０℃のアセトン５００ゴを加えて
ミキサーで１５秒間撹拌し、ブフナ一炉斗で吸引濾過す
る。これを２回繰り返し、扇風機で風乾する。このアセ
トン処理した顆粒を乳鉢で粉末にしアセトン粉末約１６
０１を得る。アセトン粉末３３．５ｇを蒸留水５００ｍ
ｊに懸濁し、室温で３０分間撹拌し、遠心して上清を採
取して粗抽出液を得る。この粗抽出液に終濃度０．０１
Ｍになるように０．２Ｍリン酸カリウム緩衝液（］：Ｈ
７，５）を添加する。０．０１Ｍリン酸カリウム緩衝液
（ＰＨ７．５）に平衡化したＤＥＡＥ−セルローズを湿
重量で約３０９加え、４℃に冷却し３０分攪拌しピロリ
ン−５−カルボン酸還元酵素を吸着させた後、ブフナ一
済斗で吸引済過し、残つた沢過残渣を０．０５Ｍおよび
０．１Ｍカリウムリン酸緩衝液（ＰＨ７．５）２００Ｗ
ＩＩに順次懸濁し、攪拌後吸引淵過し、夾雑タンパク質
を溶出させ洗浄する。Grind in a mortar until fine granules form, dry powder approx.
Get y. Add 500 g of acetone at -20°C to this, stir for 15 seconds with a mixer, and filter with suction using a Buchna furnace. Repeat this twice and air dry with a fan. Powder the acetone-treated granules in a mortar to acetone powder of about 16
Get 01. 33.5g of acetone powder in 500ml of distilled water
J, stirred at room temperature for 30 minutes, and centrifuged to collect the supernatant to obtain a crude extract. This crude extract has a final concentration of 0.01.
0.2M potassium phosphate buffer (]:H
7, 5). DEAE-cellulose equilibrated in 0.01 M potassium phosphate buffer (PH 7.5) was added in a wet weight of approximately 309 kg, cooled to 4°C and stirred for 30 minutes to adsorb pyrroline-5-carboxylic acid reductase. , suctioned and filtered with a Buchna Ikjeto, and the remaining filter residue was added to 0.05M and 0.1M potassium phosphate buffer (PH7.5) 200W.
After stirring, the mixture is successively suspended in 100% solution II and filtered through suction to elute and wash the contaminant proteins.
この残渣に０．５Ｍ塩化カリウム溶液〔０．１Ｍカリウ
ムリン酸緩衝液（ＰＨ７．５）に溶解したもの〕３００
ｍ１を加えて攪拌して酵素を溶出させ、これをブフナ一
淵斗で吸引淵過し、済液を回収する。この操作を５０ｍ
１の溶出液で２回繰り返した。Ｆ５液を合し、４Ｍ酢酸
緩衝液（ＰＨ５．５）を加えてＰＨを７．０に調節し、
１００１１１ずつ５００ｍ１ビーカ一にとつて６０℃の
熱処理を行う。熱処理後直ちに氷冷して遠心し、上清を
採取した。この上清をミリボアメンブランフイルタ一（
限外済過分子量１，０００）で約１００ｍ１まで濃縮し
た。濃縮した酵素液をウイスキングチユーブに入れ４℃
で、０．０１Ｍカリウムリン酸緩衝液（ＰＨ７．５）中
で透析して脱塩し、遠心して上清をとり、上清を同緩衝
液にて平衡化したＤＥＡＥ−セルローズカラム（３．０
×２５．０Ｃ１ｒＬ）に吸着させ、００１Ｍ，０．０５
Ｍ，０．１Ｍカリウムリン酸緩衝液（ＰＨ７．５）で順
次洗浄した。次いでＯ−０．５Ｍ塩化カリウム溶液（０
．１Ｍカリウムリン酸緩衝液に溶解させたもの）の直線
的濃度勾配をつくり酵素を溶出させた。この精製段階に
おける全活性と比活性は次の如くであり、最終生成物の
比活性は３．５であり約１５０倍精製された。To this residue, add 0.5M potassium chloride solution [dissolved in 0.1M potassium phosphate buffer (PH7.5)] 300
The enzyme is eluted by adding m1 and stirring, and this is suctioned and filtered through a Buchna Ippuchi to collect the filtrate. Repeat this operation for 50m
Repeated twice with the eluate of 1. Combine the F5 solutions, add 4M acetate buffer (PH5.5) to adjust the pH to 7.0,
Heat treatment is performed at 60° C. for each 500 ml beaker. Immediately after the heat treatment, the mixture was cooled on ice, centrifuged, and the supernatant was collected. Filter this supernatant through a millibore membrane filter (
It was concentrated to about 100 ml using ultraviolet molecular weight (extra molecular weight: 1,000). Pour the concentrated enzyme solution into a whisking tube at 4°C.
The solution was dialyzed and desalted in 0.01M potassium phosphate buffer (PH7.5), centrifuged to collect the supernatant, and the supernatant was equilibrated with the same buffer using a DEAE-cellulose column (3.0
×25.0C1rL), 001M, 0.05
Washed sequentially with M and 0.1M potassium phosphate buffer (PH7.5). Then O-0.5M potassium chloride solution (0
．． The enzyme was eluted by creating a linear concentration gradient (dissolved in 1M potassium phosphate buffer). The total activity and specific activity in this purification step were as follows, and the final product had a specific activity of 3.5 and was purified approximately 150 times.
収率は４３．５％であつた。このものは−２０℃で保存
すれば約３ケ月間安定であつた。実施例 ２
オルニチン・アミノ基転移酵素活性の測定：０．２Ｍカ
リウムリン酸緩衝液（ＰＨ７．５）２００μモル、α−
ケトグルタル酸１５μモル、ＮＡＤＨＯ．３７μモル、
Ｌ−オルニチン３０μモル、ピロリン−５−カルボン酸
還元酵素２００ミリ単位および被検試料を加えて全量を
３．０ｍ１とする。The yield was 43.5%. This product was stable for about 3 months when stored at -20°C. Example 2 Measurement of ornithine aminotransferase activity: 200 μmol of 0.2M potassium phosphate buffer (PH7.5), α-
Ketoglutaric acid 15 μmol, NADHO. 37 μmol,
Add 30 μmol of L-ornithine, 200 milliunits of pyrroline-5-carboxylic acid reductase, and the test sample to bring the total volume to 3.0 ml.
被検試料として、精製した肝オルニチン・アミノ基転移
酵素（４）とラツト肝の１０倍ホモジネート上清（Ｂ）
を用いて、その酵素活性を測定した。測定は３４０ｎｍ
における吸光度の減少を測定することにより行い、反応
の際の温度は３７℃で行つた。その結果はそれぞれ第１
図および第２図の如くであつた。実施例 ３
０．２Ｍカリウムリン酸緩衝液（ＰＨ７．５）２００μ
モル、ＮＡＤＨＯ．３７μモル、ピロリン−５−カルボ
ン酸還元酵素２００ミリ単位、精製ラツト肝オルニチン
・アミノ基転移酵素１０００ミリ単位に被検試料を加え
て全量を３ｍ１とする。As test samples, purified liver ornithine aminotransferase (4) and rat liver 10x homogenate supernatant (B)
The enzyme activity was measured using Measurement is at 340nm
The temperature during the reaction was 37°C. The results are the first
It was as shown in Fig. 2 and Fig. 2. Example 3 0.2M potassium phosphate buffer (PH7.5) 200μ
Mol, NADHO. Add the test sample to 37 μmol, 200 milliunits of pyrroline-5-carboxylic acid reductase, and 1000 milliunits of purified rat liver ornithine aminotransferase to make a total volume of 3ml.
３０℃で３０分間反応を行つたのち、３４０ｎｍにおけ
る吸光度を測定した。After reacting at 30° C. for 30 minutes, absorbance at 340 nm was measured.
その結果は第３図の如くであつた。The results were as shown in Figure 3.
Claims (1)
補酵素、ピロリン−５−カルボン酸還元酵素およびオル
ニチンまたはオルニチン・アミノ基転移酵素の何れか一
つを含む系に被検試料を加えて反応させ、還元型ピリジ
ンヌクレオチド補酵素の減少を測定することを特徴とす
るオルニチンまたはオルニチン・アミノ基転移酵素の測
定法。 ２ 還元型ピリジンヌクレオチド補酵素がニコチンアミ
ドアデニンジヌクレオチドまたはニコチンアミドアデニ
ンジヌクレオチドリン酸である特許請求の範囲第１項記
載の測定法。 ３ 還元型ピリジンヌクレオチド補酵素の減少を吸光度
によつて行う特許請求の範囲第１項記載の測定法。[Claims] 1. A test sample is added to a system containing α-ketoglutarate, reduced pyridine nucleotide coenzyme, pyrroline-5-carboxylic acid reductase, and either ornithine or ornithine aminotransferase. 1. A method for measuring ornithine or ornithine aminotransferase, which comprises reacting with pyridine and measuring the decrease in reduced pyridine nucleotide coenzyme. 2. The measuring method according to claim 1, wherein the reduced pyridine nucleotide coenzyme is nicotinamide adenine dinucleotide or nicotinamide adenine dinucleotide phosphate. 3. The measuring method according to claim 1, in which the reduction of reduced pyridine nucleotide coenzyme is measured by absorbance.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP421377A JPS5915639B2 (en) | 1977-01-18 | 1977-01-18 | Method for measuring ornithine or ornithine aminotransferase |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP421377A JPS5915639B2 (en) | 1977-01-18 | 1977-01-18 | Method for measuring ornithine or ornithine aminotransferase |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS5389493A JPS5389493A (en) | 1978-08-07 |
| JPS5915639B2 true JPS5915639B2 (en) | 1984-04-10 |
Family
ID=11578334
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP421377A Expired JPS5915639B2 (en) | 1977-01-18 | 1977-01-18 | Method for measuring ornithine or ornithine aminotransferase |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS5915639B2 (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0388637A (en) * | 1989-08-31 | 1991-04-15 | Toshiba Corp | Paper sheet conveyer |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0611238B2 (en) * | 1983-10-27 | 1994-02-16 | 有信 藤村 | Method for measuring argininosuccinate or argininosuccinate lyase |
-
1977
- 1977-01-18 JP JP421377A patent/JPS5915639B2/en not_active Expired
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0388637A (en) * | 1989-08-31 | 1991-04-15 | Toshiba Corp | Paper sheet conveyer |
Also Published As
| Publication number | Publication date |
|---|---|
| JPS5389493A (en) | 1978-08-07 |
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