Reaction #806257

ord-c9da4ef4b56a4eef9f815df5982be991

Reaction equation

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
Yield 87.4%

Reactants

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

Reagents

None

Solvents

CCCCOC(C)=O
Butyl acetate
O
water

Conditions

Detailed conditions
See reaction.notes.procedure_details.

Workup

  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
    Otherto separate
  6. 6
    Otherwas separated
  7. 7
    Washwashed with 10 mL of water
  8. 8
    ExtractionThe combined aqueous layers were extracted twice with 20 mL of butyl acetate
  9. 9
    Otherrotary evaporated under vacuum
  10. 10
    Otherto remove water
  11. 11
    workup.ADDITIONAdditional butyl acetate was added during the evaporation
  12. 12
    Otherto help remove the water
  13. 13
    OtherWhen the water was removed the butyl acetate solution
  14. 14
    Otherwas decanted from solids in the flask
  15. 15
    OtherEvaporation of the solvent under vacuum

Procedure

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.v1Patent: US07132267B2uspto-grants-2006_11