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. 1985 Apr 1;227(1):205-10.
doi: 10.1042/bj2270205.

A sensitive spectrophotometric assay for peroxisomal acyl-CoA oxidase

G M Small, K Burdett, M J Connock

A sensitive spectrophotometric assay for peroxisomal acyl-CoA oxidase

G M Small et al. Biochem J. .

Abstract

A simple spectrophotometric assay was developed for peroxisomal fatty acyl-CoA oxidase activity. The assay, based on the H2O2-dependent oxidation of leuco-dichlorofluorescein catalysed by exogenous peroxidase, is more sensitive than methods previously described. By using mouse liver samples, cofactor requirements were assessed and a linear relationship was demonstrated between dye oxidation and enzyme concentration. By using this assay on subcellular fractions, palmitoyl-CoA oxidase activity was localized for the first time in microperoxisomes of rat intestine. The assay was also adapted to measure D-amino acid oxidase activity, demonstrating the versatility of this method for measuring activity of other H2O2-producing oxidases.

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