Project information
Studying enzyme reaction rates in engineered phase separated condensates (ENZYME-LLPS tags)

Enzymes are biocatalysts at the core of many important processes in industry, medicine and research. They make a wide array of reactions possible, and with their renewable nature and scalable production are one of the cornerstones of so-called green chemistry.

Numerous biochemical reactions have been described to occur in phase-separated environments in vivo. It is hypothesized that liquid-liquid phase separation enables cells to create conditions ideal for a given reaction, increase the local concentration of reactants and to overcome kinetic limitations. Usually, the ability of a protein to phase separate is conferred by the presence of an unstructured, intrinsically disordered region. These domains can phase separate in the presence of molecular crowding agents, nucleic acids, or client proteins.

Our proposal lies in the use of phase-separating domains as fusion tags to append to relevant enzymes in order to create a new class of fusion proteins. These proteins would have the ability twould o phase separate into condensates with tremendous local enzyme concentration and, by virtue of a scaffold-client interaction, could recruit substrates for the enzyme. This lead to the rapid increase of the reaction rate, speeding up reaction times, cost savings on enzymes needed for the completion of the reaction and enabling the clustering of several enzymatic functions in one condensate. In practical terms, we envision applications in molecular diagnostics and biotechnology.