A New Spectrophotometric Assay for Enzymes of Purine Metabolism

Abstract

A new method for the determination of guanase is described. Xanthine, the product of the guanase reaction, is oxidized by xanthine oxidase, forming uric acid and hydrogen peroxide. Hydrogen peroxide is further reduced to water by catalase in the presence of ethanol. The acetaldehyde formed in this reaction step is dehydrogenated NAD or NADP dependent by aldehyde dehydrogenase. The NADH or NADPH production is measured and utilized for the calculation of the guanase activity. The sensitivity of the method can be doubled by the addition of uricase, which oxidizes uric acid to permit the formation of another mole of hydrogen peroxide.

Evaluation of the Spectrophotometric Assay of Guanase with 2,2'-Azino-di(3-ethylbenzthiazoline-6-sulphonate) (ABTS) as Chromogen

“…in humans, high äctivities have been activity can be measured via uric acid (8,9) or H 2 O 2 found in liver, kidney and brain, and little or no formed in thisreaction (10,11); O 2 consumption also activity in other organs (2,3).…”