Reaktion #56249

Reaktionsgleichung

potassium phosphate

L-lysine

acetate

3-methyl-2-benzothiazolone hydrazone hydrochloride

→

α-keto-ε -aminocaproic acid

Reaktionsbedingungen

Aufarbeitung

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

Vorschrift

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.

Quelle