L-Tyrosine → Hexyl 2-cyanoacrylate
Reaktionsgleichung
O=C[C@H]([C@@H:5]([C@@H:7]([CH2:9][OH:10])O)O)O.N[C@H](C(O)=O)C[C:14]1[CH:19]=[CH:18][C:17]([OH:20])=[CH:16][CH:15]=1.C(O)[C:25]([NH2:30])(CO)CO>>[CH3:18][CH2:19][CH2:14][CH2:15][CH2:16][CH2:17][O:20][C:9]([C:7]([C:25]#[N:30])=[CH2:5])=[O:10]Komponenten
4
Versuchsvorschrift
Overnight arabinose-induced cultures were harvested by centrifugation. The cells were resuspended in 50 mM Tris, pH 8.5. Four repeating cycles of 30 seconds sonicatation and 60 seconds rest were used to break open the cells. These samples were centrifuged to separate the soluble and insoluble fractions. The pellets were resuspended in 50 mM Tris, pH 8.5. The enzyme assay was then performed using UV grade cuvettes in a final volume of 1 mL. The assay buffer was the same 50 mM Tris at pH 8.5 used throughout the sample preparation. The substrate used was 1 mM tyrosine and the sample was 40 μg of crude cell free extract. The assay was performed at 25° C. and was run for 5 mins.