Metabolic control analysis of glycolysis

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Metabolic engineering is a tool that is increasingly used for the development of new crop varieties. The routine use of metabolic engineering in crop improvement strategies necessitates a complete understanding of mechanisms by which metabolic pathways are regulated in vivo. Glycolysis is involved in the generation of metabolic energy and in the breakdown of carbohydrates to provide carbon skeletons to a large variety of plant biosynthetic pathways. It is also involved in plant adaptations to several environmental stresses. Thus, metabolic engineering of plant glycolysis has potential applications in improvement of crops that are tailored to produce valuable natural products or to resist better to environmental stresses. However, our current knowledge on how this metabolic pathway is regulated is still incomplete. The long term objective of my research program is to better understand the regulation of plant glycolysis, and in particular how the amount of individual enzymes influences carbon flux in the glycolytic pathway. The research described in the present proposal aims at the characterization of three enzymes (hexokinase, cytosolic triosephosphate isomerase and cytosolic nucleoside diphosphate kinase) in the control over the plant glycolytic pathway. For this, we will use a multidisciplinary approach encompassing enzyme purification and characterization, manipulation of enzyme activity by transgenic means, metabolic profiling of mutants and transgenic plants and post-translational modification of proteins. The proposed research will provide training for PhD, MSc and undergraduate students. In the long term, applications of this research will allow a rationale approach to engineer carbon metabolism in plants.