An enzyme-coupled assay for acyl-CoA synthetase.
An enzyme-coupled, colorimetric method for the assay of acyl-CoA synthetase is described. Acyl-CoA formed from fatty acid and CoA by acyl-CoA synthetase was dehydrogenated by acyl-CoA oxidase. Hydrogen peroxide produced was then converted into formaldehyde in the presence of methanol by catalase. The formaldehyde reacted with a triazole compound, 4-amino-3-hydrazino-5-mercapto-1,2,4-triazole, in an alkaline condition to form a purple dye, and the absorbance was measured spectrophotometrically.
Ichihara
K
K
Department of Agricultural Chemistry, Kyoto Prefectural University, Japan.
Shibasaki
Y
Y
D003066:Coenzyme A Ligases / Q000032:analysis*; D003124:Colorimetry / Q000379:methods*; D007700:Kinetics
D003066:Coenzyme A Ligases