Genetic code expansion for biocatalysis and enzyme engineering doctoral network

The expansion of the genetic code for the incorporation of non-canonical amino acids (NCAA) in proteins provides biotechnologists with an amazing kit of novel tools. NCAA can be used in multiple applications such as investigation of the structure and dynamics of proteins, as handles for protein imaging and spectroscopy, metal chelators or as click chemistry reagents. Yet, only a few cases exist where NCAA are used in enzyme engineering to explore, improve or install new enzymatic activities. Further development has been restricted by the limited efficiency and flexibility of tools for their incorporation. This is mainly caused by the field developing separately and not being driven by desired target catalytic activities and needs of protein engineers. Therefore, this DN proposal aims to bridge this gap and bring experts from different fields together to ensure the development of the methodologies in a collaborative manner. With a threefold- training structure Learn–Teach–Create, the DN trains 10 PhDs at the boundaries of biocatalysis, synthetic biology, peptide chemistry and computational biology and brings together a team of academic and industrial partners to increase the European research capacity.

The efficiency and flexibility in NCAA incorporation are decisive for successful catalytic application; use of NCAA for selective protein conjugation facilitates application in site-selective immobilization; combination of protein folding analyses in high-throughput-screens with automatization fosters the application of NCAA in directed evolution to engineer reactivity and stability of enzymes; going a step further, NCAA can provide functional groups for designed biocatalysts for new-to-nature reactions. The proposed research affords new tools and applications, and the newly designed catalysts are envisioned to be important biotechnological processes for a greener, more environmentally friendly production of chemicals and pharmaceuticals.