Abstract

A simple method for continuous spectrophotometric assay of fructose-1,6-bisphosphate aldolase is described. The method is based on the reactivity of the product triose phosphates with cyanide to form compounds capable of the reduction of cytochrome c. At the concentrations employed, cyanide, acting catalytically, reacts with the enediol tautomer of the triose phosphates to generate the reductants, which reduce cytochrome c causing increased absorbance at 550 nm. The rate of increase in the rate of appearance of 550-nm absorbance is directly proportional to the concentration of aldolase present. The procedure is particularly useful for Class I aldolases, since the assay must be run under mildly alkaline conditions.

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