The scientific pursuits can be roughly categorized as efforts to elucidate the chemistry of potential protometabolic pathways and the synergies that arise between such chemistry and prebiotic compartments. Both tracks were highly successful revealing new facets of prebiotic chemistry. The investigated protometabolic pathways included glycolysis and the citric acid cycle, since both are believed to be ancient and the former feeds into the latter in extant biology. When run in the forward direction, the citric acid cycle extracts and captures energy from fuel sources. When run in reverse, this same cycle can be used to synthesize the building blocks of life, e.g. amino acids. For example, we found that amino acids could be produced from the ketoacids of the citric acid cycle by reductive amination with hydrogen as the electron donor and iron containing meteorites as the catalyst. Chains of amino acids, i.e. peptides, can function as effective scaffolds for the coordination of iron-sulfur clusters capable of binding substrates, features present in the contemporary citric acid cycle. Additionally, the amino acid proline was found to function as an aldol catalyst within a prebiotic triose glycolysis pathway by facilitating the reaction of glycoladlehyde-phosphate with formaldehyde to give enantioenriched glyceraldehyde-2-phosphate. NADH can be produced by the nonenzymatic reduction of NAD+ by alpha-ketoacids, and NADH can participate in the enantioselective synthesis of amino acids during nonenzymatic reductive amination. Interestingly, the production and consumption of NADH could be tied to the formation and dissipation of compartments, since NADH was found to form coacervates with positively charged polypeptides, such as poly-Arg. Lattice Model Simulations were used to assess the impact of affinity and aggregation and the impact of the micro environments within such aggregates on flux through protometabolic pathways. In addition to coacervates, the role of lipids was assessed by demonstrating how the partitioning of hemin to aggregates of single-chain amphiphiles could generate catalytic pockets with peroxidase activity. Throughout these studies, new microfluidic technologies with tunable temperature and pH were developed for the investigation of compartments and protometabolism. The completion of ProtoMet led to the awarding of 6 PhDs and one more planned shortly. One additional ESR that was brought in later and was not initially enrolled in a PhD program is now enrolled in a PhD program. All ESR went through training, had career plans, and all that have graduated have secured positions either in industry or academia. The final symposium held by the consortium was a tremendous success that brought together leading figures in the field and communicated their findings through public events.
