Electrifying Peptide Synthesis for Directed Evolution of Artificial Enzymes

Projektbeschreibung

Neuartige elektrochemische Plattformen für die Peptidsynthese ermöglichen die Entwicklung robuster Katalysatoren für künstliche Enzyme

Im gemäßigten inneren Milieu lebender Organismen wandeln die natürlichen Katalysatoren (empfindliche Proteinenzyme) kleine Moleküle für eine Vielzahl von zellulären Stoffwechselfunktionen in andere um. Viele dieser Reaktionen sind energierelevant, und ihre Nutzung zur Unterstützung von Konversionsprozessen für den Übergang zu erneuerbaren Energien würde eine nachhaltige Alternative zu den derzeit seltenen Edelmetallen darstellen. Unter den extremen Betriebsbedingungen von Brennstoffzellen oder Elektrolyseuren wird jedoch die Funktion dieser empfindlichen Enzyme beeinträchtigt. Das EU-finanzierte Projekt E-VOLUTION wird Methoden der gezielten Evolution anwenden, die bereits für natürliche Aminosäuren und biologische Bedingungen eingesetzt werden, um hochaktive und robuste künstliche Enzyme aus natürlichen und künstlichen Aminosäuren herzustellen.

Ziel

Global climate and energy challenges require efficient, robust and scalable catalysts for the conversion of renewable energies. Nature has evolved extremely active catalysts (enzymes) for the conversion of small molecules relevant to energy (H2, CO2, N2). The scalability of these enzymes offers distinct advantages over the rare, precious metals that are currently used in energy conversion. Unfortunately, the enzymes are unable to tolerate the extreme conditions of operating fuel cells or electrolyzers. Directed evolution is a powerful approach for improving enzymes, but is mostly restricted to natural amino acids and biological conditions, with limited compatibility for evolving enzymes toward enhanced resistance in abiotic systems. Here, I aim to establish directed evolution in fully abiotic systems, using artificial amino acids to make artificial enzymes that are stable even in extreme conditions. Towards this, I will establish new electrochemical peptide synthesis platforms to enable the generation of enzyme-length peptides using both natural and artificial amino acids. Extended libraries of artificial enzyme variants will be produced and screened directly on electrode microarrays. Top enzyme candidates for the conversion of H2 will be selected using fuel cell/electrolyzer conditions as the evolutionary criteria. By the end, I will have a new procedure for synthesizing libraries of full-length artificial proteins, enabling the creation of thousands of enzyme variants using artificial building blocks. The generation of high-quality datasets will be transformative to drive future machine learning-based evolution steps for both full size enzymes and small-molecule catalysts with applications beyond H2 evolution. We will have discovered highly active catalysts able to sustain conditions of large-scale energy conversion devices, accelerating breakthroughs toward the economically competitive use of renewable energies for fuel and chemical production.