Biological Chemistry: Directed Evolution of Proteins

Abstract

During a four-week block course in the semester break, we will carry out biological-chemical enzyme evolution experiments using molecular genetic mutation technologies and in vivo selection in recombinant bacterial strains. The class with its very dense program consists of the practical course itself and an integrated series of seminar/lecture sessions.

Objective

All technologies used for the experiments will be explained to the students in theory and in practice with the goal that they will be able to independently apply them in the last week of the course. After the course, an individual report about the results obtained has to be prepared.

Content

We intend to carry out biological-chemical enzyme evolution experiments using molecular genetic mutation technologies and in vivo selection in recombinant bacterial strains. By working in parallel, up to 9 student teams composed of 2 participants each should evolve a variety of different variants of a catalyst. Individual proteins will be purified and subsequently characterized using several different spectroscopic methods. The detailed chemical-physical analyses include determination of the enzymes' kinetic parameters and the integrity of the secondary structure. The results obtained from the individual evolution experiments will be assembled, compared and discussed at the end of the class in a final seminar. We expect that during this lab course we will not only generate novel enzymes, but also gain new insights into the reaction mechanism of the investigated catalysts.

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