Reaction #829366

ord-9f3d0e9b6f1948cfbc2e6b3a241cc485

Reaction equation

O=C(O)C1=NC(C(=O)O)CC=C1
dihydrodipicolinate
NC(=O)C1=CN([C@@H]2O[C@H](COP(=O)(O)OP(=O)(O)OC[C@H]3O[C@@H](n4cnc5c(N)ncnc54)[C@H](OP(=O)(O)O)[C@@H]3O)[C@@H](O)[C@H]2O)C=CC1
NADPH
O=C(O)C1=NC(C(=O)O)CC=C1
Dihydrodipicolinate
→
O=C(O)C1=NC(C(=O)O)CC=C1
dihydrodipicolinate
NC(=O)C1=CN([C@@H]2O[C@H](COP(=O)(O)OP(=O)(O)OC[C@H]3O[C@@H](n4cnc5c(N)ncnc54)[C@H](OP(=O)(O)O)[C@@H]3O)[C@@H](O)[C@H]2O)C=CC1
NADPH
O=C([O-])c1cccc(C(=O)[O-])n1
PDC

Reactants

O=C(O)C1=NC(C(=O)O)CC=C1
dihydrodipicolinate
NC(=O)C1=CN([C@@H]2O[C@H](COP(=O)(O)OP(=O)(O)OC[C@H]3O[C@@H](n4cnc5c(N)ncnc54)[C@H](OP(=O)(O)O)[C@@H]3O)[C@@H](O)[C@H]2O)C=CC1
NADPH
O=C(O)C1=NC(C(=O)O)CC=C1
Dihydrodipicolinate

Reagents

None

Conditions

Detailed conditions
See reaction.notes.procedure_details.

Workup

  1. 1
    Othercan be produced

Procedure

Dihydrodipicolinate reductase ternary crystals can be produced by mixing dihydrodipicolinate reductase with a 1.5 molar excess of NADPH and a 2.5 molar excess of 2,6-pyridinedicarboxylate (PDC) for 30-45 minutes at 4° C. to obtain a ternary complex of NADPH, PDC and dihydrodipicolinate reductase. Next, a precipitant solution containing polyethylene glycol, a salt and a buffer is provided. A droplet of dihydrodipicolinate reductase solution is mixed with a droplet of precipitant solution to obtain a mixed droplet solution. The mixed droplet solution is suspended over a well of precipitant solution in a sealed container. The vapor pressure of the precipitant solution in the well must be lower than the vapor pressure of the mixed droplet solution. The mixed droplet solution is allowed to stand for a period of time sufficient for dihydrodipicolinate reductase ternary crystals to grow to a predetermined size. Crystals appear in one to three weeks.

Source

DOI: 10.6084/m9.figshare.5104873.v1Patent: US05554522uspto-grants-1996_09