反応式
P([O-])([O-])([O-])=O.[K+].[K+].[K+].N[C@H:10]([C:16]([OH:18])=[O:17])[CH2:11][CH2:12][CH2:13][CH2:14][NH2:15].C([O-])(=[O:21])C.Cl.CN1C2C=CC=CC=2SC1=NN>>[O:21]=[C:10]([CH2:11][CH2:12][CH2:13][CH2:14][NH2:15])[C:16]([OH:18])=[O:17]参加物
実験手順
The activity of the present enzyme was measured in accordance with Soda's method (Analytical Biochemistry Vol. 25, p. 228, 1968) in the following manner. A reaction mixture consisting of 0.7 ml of 0.1 M potassium phosphate buffer (pH 8.0), 0.1 ml of catalase (750 U/ml), 0.1 ml of 0.1 M L-lysine solution and 0.1 ml of the present enzyme solution was incubated at 37° C. for 20 minutes with gentle shaking. After the incubation the reaction was terminated by the addition of 0.1 ml of 25% trichloroacetic acid. To the resulting reaction mixture were added 1.9 ml of 1 M acetate buffer (pH 5.0) and 0.8 ml of 0.1% 3-methyl-2-benzothiazolone hydrazone hydrochloride solution. The mixture was further incubated at 30° C. for 30 minutes and then allowed to cool to room temperature, after which measurement of optical density at 318 nm was carried out. The formed α-keto-ε -aminocaproic acid thus formed was determined from the resulting calibration curve. One unit of the enzyme was defined as the quantity of the enzyme catalyzing the formation of 1 μmol of α-keto-ε-aminocaproic acid at 37° C. per minute. The relative activity of the enzyme was also assayed by manometric and polarographic determination of the oxygen consumption.
後処理
- customAfter the incubation the reaction was terminated by the addition of 0.1 ml of 25% trichloroacetic acid
- customTo the resulting reaction mixture
- waitThe mixture was further incubated at 30° C. for 30 minutes
出典と根拠
- ORDレコード
- ord-cf1a7cc260784b448642a5e80cbf8003
- 出典データセット
- uspto-grants-1980_11
- 特許番号
- US04234691
関連ノート
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