Réaction #1244288

Équation de réaction

ethyl 4-chloroacetoacetate

glucose

2

Glucose

potassium phosphate

NaCl

glucose

Na2CO3

NADP

→

ethyl (S)-4-chloro-3-hydroxybutyrate

Rendement 97.0%

Solvants

butyl acetate

butyl acetate

Conditions de réaction

Traitement

1. 1
   workup.ADDITIONTo this was added ketoreductase SEQ ID NO
2. 2
   workup.DISSOLUTIONOnce dissolved
3. 3
   Autreto form an emulsion
4. 4
   TempératureThe pH was maintained between 6.8 and 7 by an automatic titrater that
5. 5
   workup.ADDITIONAfter 40 hours the automated addition of the base
6. 6
   AutreThe layers were separated
7. 7
   Lavagethe aqueous phase was washed with ethyl acetate (500 mL)
8. 8
   SéchageThe combined organics were dried over anhydrous sodium sulfate
9. 9
   Filtrationfiltered
10. 10
    Autreevaporated on a rotary evaporator

Mode opératoire

To a well-stirred solution of 100 mM potassium phosphate buffer, 500 mM NaCl, pH 7 (1 L) at room temperature was added glucose (160 g, 830 mmoles, 1.1 equiv). To this was added ketoreductase SEQ ID NO: 2 (0.9 g), glucose dehydrogenase-S06 SEQ ID NO: 10 (0.5 g) and NADP (0.5 g) as lyophilized powders. Once dissolved, butyl acetate (500 mL) was added to form an emulsion. To this emulsion was added a solution of ethyl 4-chloroacetoacetate (10 g, 608 mmoles) in butyl acetate (500 mL), dropwise over 3 hours. The pH was maintained between 6.8 and 7 by an automatic titrater that dispensed Na2CO3 (2M in water, about 160 mL total). After 40 hours the automated addition of the base had ceased and there was no residual starting material by gas chromatography. The layers were separated, and the aqueous phase was washed with ethyl acetate (500 mL). The combined organics were dried over anhydrous sodium sulfate, filtered and evaporated on a rotary evaporator, to give essentially pure (˜97%) ethyl (S)-4-chloro-3-hydroxybutyrate.

Source