Biological Chemistry A: Technologies for Directed Evolution of Enzymes

Abstract

During this course, methodologies will be taught for biological-chemical enzyme evolution experiments using molecular genetic mutation technologies and in vivo selection in recombinant bacterial strains. The class consists of afternoon sessions and involves both practical work to generate the biomaterials and data needed for a research project 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 for the course project and in future research endeavors. After the course, an individual report about the results obtained has to be prepared.

Content

This class conducts and supports experiments for a specifically designed genuine research project. We will carry out biological-chemical enzyme evolution experiments using molecular genetic mutation technologies and in vivo selection in recombinant bacterial strains. As a focus of this course, all relevant technologies will be taught to the students, such as the preparation of competent cells, production and isolation of DNA fragments, transformation of gene libraries, and DNA sequencing. In addition, novel enzyme catalysts will be generated, purified, and 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. 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 catalyst.

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