Measurement of Enzyme Activities and Optimization of Continuous and Discontinuous Assays
Langue
en
Chapitre d'ouvrage
Ce document a été publié dans
Current Protocols in Plant Biology, Current Protocols in Plant Biology. 2016p. np
John Wiley and Sons Inc
Résumé en anglais
The measurement of enzyme activities represents an important step towards the understanding of biological networks. Continuous or discontinuous assays can be used, as well as highly sensitive assays, depending on the ...Lire la suite >
The measurement of enzyme activities represents an important step towards the understanding of biological networks. Continuous or discontinuous assays can be used, as well as highly sensitive assays, depending on the abundance of the enzymes under study. To exemplify such methods, two protocols for phosphoenolpyruvate carboxylase activity (EC 4.1.1.31) in plant extracts are given. For this, an extraction protocol is also described. Then, an optimization protocol for enzyme assays using enzymatic, chemical, or biological standards is proposed. This protocol evaluates in one run the optimal extract dilution, the recovery of a standard, and the technical error in a given matrix. The interest of using biological standard in routine measurements is highlighted.< Réduire
Mots clés
métabolisme
physiologie végétale
Mots clés en anglais
cycling assay
discontinuous assay
enzyme activity
optimization
PEPC
recovery
continuous assay
Project ANR
Centre français de phénomique végétale - ANR-11-INBS-0012
Origine
Importé de halUnités de recherche