Guanine Aminohydrolase 활성의 신비색측정법

Abstract

A new colorimetric assay method for guanine deaminase was reported, coupling the enzyme with xanthine oxidase and formazan reaction system adding phenazine methosulfate and INT (2-(p-iodophenyl)-3-(p-nitrophemyl) -5-phenyl tetrazolium chloride). The method was proved to be applicable both for the assay of GOA activity and for visualization of the enzymogram after electrophoresis. The assay system consisted of GDA, XO as much as 20 fold of GOA, O. 161'mole of PMS, and 1. 51' mole of INT in a final volume of 3.0 ml. Tris buffer, pH 8. O. The reaction was started by the addition of 1. 5 I' mole guanine, followed by 30 minutes incubation at 30°C. The present data was inconsistent with the reported data, but could be converted by a factor for the purpose of comparison. It appears to be a good merit to apply the present method in estimating the GOA activity after staining electrophorogram with the formazan reaction introduced in the present study directly utilizing extract of visual enzyme band.