Optimization of the performance of an artificial imine reductase by directed evolution and encapsulation

The present PhD dissertation summarizes the research findings of the research group of prof. Dr. Thomas R. Ward from the period of September 2013 to December 2017.
The main goal of the Ward group is the development and optimization of artificial metalloenzymes for their use in asymmetric catalysis. These hybrid catalysts, resulting from an incorporation of a metal–containing cofactor within a protein or DNA scaffold, can be optimized by either chemical or genetic means.
The present work aims at the genetic optimization of the protein scaffold and exploration of the use of the resulting hybrid catalysts in nano–applications. Apart from a general introduction section, this dissertation is divided into two chapters. Each chapter represents research performed in context of several projects, which were published or will be submitted to peer-reviewed journals. These chapters contain a brief introduction into the topic, followed by the corresponding publications, and author contributions. The electronic supplementary information for each publication can be found in the Appendices chapter.
The first chapter describes our efforts in genetic optimization of artificial metalloenzymes, design of a screening library and directed evolution of the resulting mutants. The second chapter illustrates two possibilities of applying artificial metalloenzymes in nano-applications, specifically the immobilization on silica nanoparticles or encapsulation in a ferritin protein cage.