An HPLC method for the assay of UDP-glucose pyrophosphorylase and UDP-glucose-4-epimerase in Solieria chordalis (Rhodophyceae)

• Published in: Phytochemical analysis 2001-11, Vol.12 (6), p.363-365
• Main Authors: Goulard, F., Diouris, M., Deslandes, E., Floc'h, J. Y.
• Format: Article
• Language: English
• Subjects: Chromatography, High Pressure Liquid - methods; High-performance liquid chromatography; Marine; Rhodophyta; Rhodophyta - enzymology; Solieria chordalis; Spectrum Analysis; UDP-glucose pyrophosphorylase; UDP-glucose-4-epimerase; UDPglucose 4-Epimerase - isolation & purification; UDPglucose 4-Epimerase - metabolism; Uridine Diphosphate Galactose - metabolism; Uridine Diphosphate Glucose - metabolism; Uridine Triphosphate - metabolism; UTP; UTP-Glucose-1-Phosphate Uridylyltransferase - isolation & purification; UTP-Glucose-1-Phosphate Uridylyltransferase - metabolism
• ISSN: 0958-0344; 1099-1565
• DOI: 10.1002/pca.604

An efficient method to assay both UDP‐glucose pyrophosphorylase and UDP‐glucose‐4‐epimerase in a crude extract of the red seaweed, Solieria chordalis is described. The method is based on the direct quantification by reverse‐phase high‐performance liquid chromatography of the UDP‐sugars generated in the reaction mixture. UDP‐glucose, UDP‐galactose and UTP were detected by spectrophotometry at 254 nm and their recoveries ranged from 97 to 100%. In the course of the reaction, a correlation was observed between the reduction in the area of the substrate peak and the occurrence of product peak(s). This highly reproducible method for enzyme assay is fast since no intermediate reaction mixture is required. Copyright © 2001 John Wiley & Sons, Ltd.