Analysis of enzyme activities

Guillaume G. Menard (1) , Benoit Biais (1) , Duyên D. Prodhomme (1) , Patricia Ballias (1) , Yves Y. Gibon (1)

1 BFP - Biologie du fruit et pathologie

The evaluation of enzyme activities, especially their capacities, represents an important step towards the modelling of biochemical pathways in living organisms. The implementation of microplate technology enables the determination of up to >50 enzymes in relatively large numbers of samples and in various biological materials. Most of these enzymes are involved in central metabolism and several pathways are entirely covered. Direct or indirect assays can be used, as well as highly sensitive assays, depending on the abundance of the enzymes under study. To exemplify such methods, protocols for UDP-glucose pyrophosphorylase (E.C. 2.7.7.9) operating in real time and for pyrophosphate:fructose-6-phosphate 1-phosphotransferase (E.C. 2.7.1.90) are presented.

Mots clés: enzyme activity; direct assay; stopped assay; cycling assay

Mots clés: microplate

Domaines: Biologie végétale

Guillaume G. Menard, Benoit Biais, Duyên D. Prodhomme, Patricia Ballias, Yves Y. Gibon. Analysis of enzyme activities. Plant Metabolic Flux Analysis : Methods and Protocols, 1090, Springer Science+Business Media, 2014, Methods in Molecular Biology, 978-1-62703-687-0. ⟨10.1007/978-1-62703-688-7_15⟩. ⟨hal-02799530⟩