529-0739-01L Biological Chemistry B: New Enzymes from Directed Evolution Experiments
|Autumn Semester 2018
|P. A. Kast
|yearly recurring course
|Language of instruction
|Number of participants limited to 12.
|During the block course in the fall semester, 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.
|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.
|The class deals with a specifically designed and genuine research project. 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, teams of 2 participants each will generate a variety of different variants of a chorismate mutase. Individual enzyme catalysts will be purified and subsequently characterized using several different spectroscopic methods. The detailed chemical-physical analyses include determination of the enzymes' kinetic parameters, their molecular mass, and the integrity of the protein structure. The results obtained from the individual evolution experiments will be 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 mechanistic insights into the investigated catalyst.
|A script will be distributed to the participants on the first day of the course.
|General literature to "Directed Evolution" and chorismate mutases, e.g.:
– Taylor, S. V., P. Kast & D. Hilvert. 2001. Investigating and engineering enzymes by genetic selection. Angew. Chem. Int. Ed. 40: 3310-3335.
– Jäckel, C., P. Kast & D. Hilvert. 2008. Protein design by directed evolution. Annu. Rev. Biophys. 37: 153-173.
– Roderer, K. & P. Kast. 2009. Evolutionary cycles for pericyclic reactions – Or why we keep mutating mutases. Chimia 63: 313-317.
Further literature will be indicated in the distributed script.
|Prerequisites / Notice
|This laboratory course will involve experiments that require a tight schedule and, particularly in the second half, very long (!) working days. The maximum number of participants for the laboratory class is limited, but surplus applicants may contact P. Kast directly to have their names added to a waiting list. A valid registration is considered a commitment for attendance of the entire course, as involved material orders and experimental preparations are necessary and, once the class has started, the flow of the experiments must not be interrupted by individual absences. In case of an emergency, please immediately notify P. Kast. For more information see Link, from where you can also download a flyer.