An HPLC method for the assay of UDP-glucose pyrophosphorylase and UDP-glucose-4-epimerase in Solieria chordalis (Rhodophyceae).
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.
Goulard
F
F
Laboratoire d'Ecophysiologie et de Biotechnologies des Algues Marines, IUEM-UBO, Technopole Brest-Iroise, place Nicolas Copernic, 29280 Plouzane, France. F.Goulard@univ-brest.fr
Diouris
M
M
Deslandes
E
E
Floc'h
J Y
JY
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D000461:Rhodophyta / Q000201:enzymology*; D013057:Spectrum Analysis; D014534:UDPglucose 4-Epimerase / Q000302:isolation & purification* / Q000378:metabolism; D005957:UTP-Glucose-1-Phosphate Uridylyltransferase / Q000302:isolation & purification* / Q000378:metabolism; D014531:Uridine Diphosphate Galactose / Q000378:metabolism; D014532:Uridine Diphosphate Glucose / Q000378:metabolism; D014544:Uridine Triphosphate / Q000378:metabolism
D014531:Uridine Diphosphate Galactose; D005957:UTP-Glucose-1-Phosphate Uridylyltransferase; D014534:UDPglucose 4-Epimerase; D014544:Uridine Triphosphate; D014532:Uridine Diphosphate Glucose