Reaction #1673829

ord-980771627a7b408582d974db79e9a5c9

Reaction equation

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
CCOC(=O)C1CCCC1=O
ethyl 2-oxocyclopentanecarboxylate
O=P([O-])([O-])[O-]
phosphate
O=C[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO
glucose
NC(=O)c1ccc[n+]([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)c1
NADP+
CCOC(=O)C1CCCC1=O
ethyl 2-oxocyclopentanecarboxylate
O=C[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO
glucose
O=C[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO
glucose
→
CCOC(=O)C1CCCC1O
ethyl 2-hydroxycyclopentane-carboxylate

Reactants

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
CCOC(=O)C1CCCC1=O
ethyl 2-oxocyclopentanecarboxylate
O=P([O-])([O-])[O-]
phosphate
O=C[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO
glucose
—
reaction mixture
—
( 1 )
NC(=O)c1ccc[n+]([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)c1
NADP+
CCOC(=O)C1CCCC1=O
ethyl 2-oxocyclopentanecarboxylate
—
1U
O=C[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO
glucose
—
2U
—
( 1 )
O=C[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO
glucose

Reagents

None

Conditions

Detailed conditions
See reaction.notes.procedure_details.

Workup

  1. 1
    Otherper 1 minute
  2. 2
    Otherwas incubated at 30° C.
  3. 3
    Otherthe enzymatic reaction
  4. 4
    Extractionthe reaction mixture was extracted
  5. 5
    ExtractionOn the other hand, the extract
  6. 6
    workup.DISTILLATIONabove was distilled off

Procedure

The asymmetric reduction of ethyl 2-oxocyclopentanecarboxylate was carried out by using this reductase (1). Namely, 4 ml of the reaction mixture containing the reductase (1) (2U), NADPH (0.8 μmol), ethyl 2-oxocyclopentanecarboxylate (100 μmol, 16 mg), phosphate buffer solution (pH 7.0, 400 μmol), glucose dehydrogenase [manufactured by Sigma, Co., U.S.A.; 2U (wherein 1U is defined as the amount of the enzyme which reduces 1 μmol of NADP+ per 1 minute in the presence of glucose)] and glucose (100 μmol) was incubated at 30° C. with gentle shaking to promote the enzymatic reaction. After 6 hour-incubation, the reaction mixture was extracted according to the same manner as in Example 3. The extract was analyzed by gas chromatography described in Example 1. As a result, the yield of the cis isomer was 15.9 mg (reduction yield >99%), and neither the remaining substrate nor the trans isomer was detected. On the other hand, the extract above was distilled off according to the the same manner as in Example 3 to obtain the reduction product, ethyl 2-hydroxycyclopentane-carboxylate. The resulting reduction product was analyzed by high performance liquid chromatography as described in Example 1. As a result, only the cis 2 isomer was detected and no other isomers were detected.

Source

DOI: 10.6084/m9.figshare.5104873.v1Patent: US05215919uspto-grants-1993_06