Abstract
The reaction mixture contained, in a total volume of 1.00m1, enzyme, p-coumaric acid (2.5pmol), dihydroxyfumaric acid (30pmol), KH2P04 (8.3pmol) and sufficient KOH to adjust the pH to 6. Incubations were carried out for 0.5h at 25°C and caffeic acid production was assayed as described by Halliwell(l975). 100% corresponded to a rate of 60nmol of caffeic acid formed in 0.5h. Oxidation of dihydroxyfumarate was followed by the decrease in E300 (althoughp-coumaric acid also absorbs light at 300nm, very little is used up during a 30min incubation). Samples (10~1) were removed from the reaction mixture at intervals, added to lOml of water and the absorbance recorded at once. Units of superoxide dismutase were as described by McCord & Fridovich (1969).
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