Abstract
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.
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Notes
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Conflict of Interest The authors declare that they have no conflict of interest.
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Financial support from INRA, Région Aquitaine, Eranet KBBE SAFQIM, Eranet Erasysbio+ FRIM, FP7 KBBE DROPS and SFR BIE is gratefully acknowledged.
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Ménard, G., Biais, B., Prodhomme, D., Ballias, P., Gibon, Y. (2014). Analysis of Enzyme Activities. In: Dieuaide-Noubhani, M., Alonso, A. (eds) Plant Metabolic Flux Analysis. Methods in Molecular Biology, vol 1090. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-688-7_15
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DOI: https://doi.org/10.1007/978-1-62703-688-7_15
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