反応 #1244288

反応方程式

ethyl 4-chloroacetoacetate

glucose

2

Glucose

potassium phosphate

NaCl

glucose

Na2CO3

NADP

→

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

収率 97.0%

溶媒

butyl acetate

butyl acetate

反応条件

後処理

1. 1
   workup.ADDITIONTo this was added ketoreductase SEQ ID NO
2. 2
   workup.DISSOLUTIONOnce dissolved
3. 3
   その他to form an emulsion
4. 4
   温度The 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
   その他The layers were separated
7. 7
   洗浄the aqueous phase was washed with ethyl acetate (500 mL)
8. 8
   乾燥The combined organics were dried over anhydrous sodium sulfate
9. 9
   ろ過filtered
10. 10
    その他evaporated on a rotary evaporator

実験手順

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.

出典