反応 #747440

反応方程式

pyruvate

formate

Lactate

formate

5,5,5-trifluoro-2-oxopentanoic acid

D,L-alanine

pyridoxal phosphate

sodium formate

NAD

lactate

NADH

NADH

→

(R)-5,5,5-trifluoro-2-aminopentanoic acid

溶媒

potassium phosphate

反応条件

後処理

1. 1
   温度The yields were increased
2. 2
   workup.ADDITIONby adding auxiliary enzymes
3. 3
   その他Reaction

実験手順

The yields were increased by adding auxiliary enzymes to reduce pyruvate to lactate. Lactate dehydrogenase requires NADH as a cofactor. The NADH was regenerated using formate dehydrogenase. A solution containing 5,5,5-trifluoro-2-oxopentanoic acid (100 mg, 0.588 mmoles), D,L-alanine (200 mg, 2.244 mmoles), 0.02 mM pyridoxal phosphate, sodium formate (68 mg, 1 mmole), NAD (3.31 mg, 5 μmoles) L-lactate dehydrogenase cloned from rabbit muscle (Biocatalytics LDH-103, 0.107 mg, 15 units), and formate dehydrogenase (0.5 mL, 15 units cloned from Pichia pastoris and expressed in Escherichia coli) in 0.1 M potassium phosphate buffer, pH 7.5, was incubated with R-transaminase AT-103 from Biocatalytics (5 mg, 44units) or cloned R-transaminase from Bacillus thuringiensis SC16569 (0.5 mL, 10 units) at 30° C. in a total volume of 5 mL in 15 mL tubes. Reaction yields of (R)-5,5,5-trifluoro-2-aminopentanoic acid of 94% and 91% were obtained with AT-103 and BMS transaminases, respectively. Ee was 100% in both cases.

出典