Reaction #58480

ord-8c87408e6e1e4db5a86984764b0046b0

Reaction equation

CCC(CC)COC(C(=O)N(C)CC[NH+](C)C)(c1ccccc1)c1ccccc1.[Cl-]
X-100
O=C1CC(C(=O)[O-])NC(=O)N1
dihydroorotate
O=C1C=CC(=Nc2cc(Cl)c(O)c(Cl)c2)C=C1
2,6-dichloroindophenol
O=C1CC(C(=O)[O-])NC(=O)N1
dihydroorotate
[Cl-].[K+]
KCl
→
O=C(O)C1=C(C(=O)O)COC1
2,5-dihydro furan 3,4-dicarboxylic acid

Reactants

CCC(CC)COC(C(=O)N(C)CC[NH+](C)C)(c1ccccc1)c1ccccc1.[Cl-]
X-100
O=C1CC(C(=O)[O-])NC(=O)N1
dihydroorotate
O=C1C=CC(=Nc2cc(Cl)c(O)c(Cl)c2)C=C1
2,6-dichloroindophenol
O=C1CC(C(=O)[O-])NC(=O)N1
dihydroorotate
[Cl-].[K+]
KCl

Reagents

None

Solvents

Cl.NC(CO)(CO)CO
TrisHCl

Conditions

Detailed conditions
See reaction.notes.procedure_details.

Workup

  1. 1
    Otherat 30° C.
  2. 2
    workup.ADDITIONThe components were mixed
  3. 3
    OtherThe course of reaction
  4. 4
    Otherin absorption at 600 nm for 2 min.

Procedure

The standard assay mixture contained 50 μM decyclo ubichinone, 100 μM dihydroorotate, 60 μM 2,6-dichloroindophenol, as well as 20 mU DHODH. The volume activity of the recombinant enzyme used was 30 U/ml. Measurements were conducted in 50 mM TrisHCl (150 mM KCl, 0,1% Triton X-100, pH. 8,0) at 30° C. in a final volume of 1 ml. The components were mixed, and the reaction was started by adding dihydroorotate. The course of reaction was followed by spectrophotometrically measuring the decrease in absorption at 600 nm for 2 min.

Source

DOI: 10.6084/m9.figshare.5104873.v1Patent: US07423057B2uspto-grants-2008_09