A Rapid and Sensitive Spectrophotometric Method for Assaying Polygalacturonase Using 2-Cyanoacetamide1

Open Access
Publisher Website
Google Scholar
Abstract
A new assay for nanomole amounts of reducing sugar using 2-cyanoacetamide has been modified and adapted to assay for exo- and endo-polygalacturonase in fruit extracts of tomato (Lycopersicon esculentum Mill. cv. Heinz 1350) with increased sensitivity (140%) and simplicity over currently used methods. Linearity was observed with galacturonic acid as a standard up to 250 nmol; the lower limit of detection was 1 nmol. Polygalacturonase product formation was linear for 3 hours and was proportional to the amount of enzyme in the reaction. Thin-layer chromatography of reaction products revealed a range of uronic acid oligosaccharides as well as galacturonic acid. Thus, both exo- and endo-polygalacturonase were active in the extracts.

This publication has 0 references indexed in Scilit: