Project 2: Recycling of molecular energy carriers in a synthetic cell
Our research group is embedded in the nationwide research programme BaSyC (Building a Synthetic Cell) of the Netherlands. Therein we mainly work on the integration of synthetic metabolic pathways responsible for energy provision and cellular homeostasis in the synthetic cell. (1) Recently our group developed a chemically fueled enzymatic metabolic pathway to regenerate ATP from ADP in vesicular systems.(2) The pathway demonstrates metabolic energy conservation and basic physicochemical homeostasis in a minimal cell-like system. The next step towards a fully autonomous synthetic cell metabolism will be to extend the enzymatic metabolic reaction network to (re)generate also other important energy carriers, which are needed to fuel and sustain the full machinery of cellular non-equilibrium processes.
Hence, the aim of this research project is to enzymatically recycle various nucleotide species through the energy provided by ATP. The research project starts with characterizing the single enzymatic reactions needed for energy regeneration and then combining those to an additional reaction module. Afterwards, the new enzymatic reaction module is coupled to the established ATP regeneration pathway in vesicles. This research project will be complemented by establishing specific molecular sensors and assay reactions for the detection of different nucleotide species in an integrated system. Within this research project, you will familiarize with the design of kinetic assays, cascaded enzymatic reaction networks, reconstitution of complex enzyme systems in vesicular systems as nanoreactors.
(1) H. R. Sikkema, B. F. Gaastra, T. Pols, B. Poolman, ChemBioChem. 2019, 20, 2581-2592
(2) T. Pols et al., Nat. Commun., 2019, 10, 4239