Réaction #806257

ord-c9da4ef4b56a4eef9f815df5982be991

Équation de réaction

CCOC(=O)CC(=O)CCl
Ethyl 4-chloroacetoacetate
O=C[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO
glucose
[Na+].[OH-]
NaOH
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)C[C@H](O)CCl
ethyl (S)-4-chloro-3-hydroxybutyrate
Rendement 87.4%

Réactifs

CCOC(=O)CC(=O)CCl
Ethyl 4-chloroacetoacetate
O=C[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO
glucose
—
42
[Na+].[OH-]
NaOH
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

Réactifs

Aucun

Solvants

CCCCOC(C)=O
Butyl acetate
O
water

Conditions de réaction

Conditions détaillées
See reaction.notes.procedure_details.

Traitement

  1. 1
    workup.ADDITIONTo a 100 mL vessel connected to an automatic titrater by a pH electrode and a feeding tube for addition of base
  2. 2
    workup.ADDITIONTo this solution was charged ketoreductase SEQ ID NO
  3. 3
    workup.ADDITION1 mL was charged rapidly
  4. 4
    workup.ADDITIONthe remainder was then charged at a rate of 1 mL/hr)
  5. 5
    Autreto separate
  6. 6
    Autrewas separated
  7. 7
    Lavagewashed with 10 mL of water
  8. 8
    ExtractionThe combined aqueous layers were extracted twice with 20 mL of butyl acetate
  9. 9
    Autrerotary evaporated under vacuum
  10. 10
    Autreto remove water
  11. 11
    workup.ADDITIONAdditional butyl acetate was added during the evaporation
  12. 12
    Autreto help remove the water
  13. 13
    AutreWhen the water was removed the butyl acetate solution
  14. 14
    Autrewas decanted from solids in the flask
  15. 15
    AutreEvaporation of the solvent under vacuum

Mode opératoire

To a 100 mL vessel connected to an automatic titrater by a pH electrode and a feeding tube for addition of base was charged a solution of glucose (12. g) in water (30 mL). To this solution was charged ketoreductase SEQ ID NO: 42 (100 mg); 50 mg GDH SEQ ID NO: 66 and NADP (6.25 mg). Butyl acetate (10 ml) was then charged. Ethyl 4-chloroacetoacetate (10 g) was then charged via syringe pump as follows: 1 mL was charged rapidly and the remainder was then charged at a rate of 1 mL/hr). The pH was maintained at 7 by the automatic titrater by the addition of 4M NaOH over 18 hours hrs. The stirring was stopped and the phases allowed to separate. The organic layer included some emulsion. The organic layer, including some emulsion, was separated and washed with 10 mL of water. The combined aqueous layers were extracted twice with 20 mL of butyl acetate. The organic extracts were combined and rotary evaporated under vacuum to remove water. Additional butyl acetate was added during the evaporation to help remove the water. When the water was removed the butyl acetate solution was decanted from solids in the flask. Evaporation of the solvent under vacuum then gave 8.85 g of ethyl (S)-4-chloro-3-hydroxybutyrate (87.4% yield) of very good purity.

Source

DOI: 10.6084/m9.figshare.5104873.v1Brevet: US07132267B2uspto-grants-2006_11