Abstract
A relatively simple assay procedure for measuring the reactions catalyzed by polyuronic acid C-5 epimerases has been developed. Action of C-5 epimerases inverts the C-6 carboxyl group of polyuronic acids converting β-linked residues into α-linked residues or vice versa. The assay takes advantage of the greater susceptibility of the acid hydrolysis of α-glycosidic linkages than β-glycosidic linkages. The method involves the partial acid hydrolysis of the polyuronic acid before and after reaction with the C-5 epimerase. The greater or lesser amounts of uronic acid released (solubilized) before and after reaction of the C-5 epimerase are a measure of the amount of α- or β-glycosidic linkages that are formed and a measure of the amount of catalysis by the enzyme.
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