反応 #470401

反応方程式

methanol

NaOH

4-(4-nitrobenzyl)pyridine

NADPH

→

4-(1-Nitrobenzyl)pyridine

溶媒

potassium phosphate

ethylacetate acetone

反応条件

後処理

1. 1
   その他A typical reaction
2. 2
   その他the vial was sealed
3. 3
   workup.WAITThe reaction was incubated at 80° C. for 20 minutes
4. 4
   温度chilled on ice
5. 5
   workup.ADDITIONThe solution was mixed thoroughly

実験手順

A typical reaction contained enzyme (1.0 ml, 1×10−6 M) in potassium phosphate buffer (0.1 M, pH 8.0), alkene (1.0×10−3 M), and methanol (1%) in a vial. The reaction was initiated with NADPH (50 μl, 1.0×10−3 M), and allowed to stir aerobically for 5 minutes. 300 μl of a stock solution of 4-(4-nitrobenzyl)pyridine (5% w/w in acetone) was added to the reaction, and the vial was sealed. The reaction was incubated at 80° C. for 20 minutes and chilled on ice. 600 μl of an ethylacetate/acetone (5:2) solution and 300 μl 5 M aqueous NaOH was added to the reaction solution. The solution was mixed thoroughly, and the absorbance of the organic layer was measured at 540 nm in a glass 96-well plate with microplate spectrophotometer.

出典