反应详情

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.

1. customA typical reaction
2. customthe vial was sealed
3. waitThe reaction was incubated at 80° C. for 20 minutes
4. temperaturechilled on ice
5. additionThe solution was mixed thoroughly