Acetate → 6-Amino-2-oxohexanoic acid

HRID 27971

Reaktionsgleichung

Reaktionsgleichung von HRID 27971
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]

Komponenten

5

Versuchsvorschrift

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.

Aufarbeitung

  1. customAfter the incubation the reaction was terminated by the addition of 0.1 ml of 25% trichloroacetic acid
  2. customTo the resulting reaction mixture
  3. waitThe mixture was further incubated at 30° C. for 30 minutes

Quellen und Nachweise

ORD-Eintrag
ord-9791e86138304557a7ae11284cfb639a
Quelldatensatz
uspto-grants-1980_09
Patentnummer
US04224407

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