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529-0739-10L 13 Credits MSC D-CHAB
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Biological Chemistry A: Technologies for Directed Evolution of Enzymes

Lecturers & Examiners: Prof. Dr. Peter Andreas Kast
Advanced laboratory course or internship depending on lab course Biological Chemistry B Candidates must inquire with P. Kast no later than September 1st whether course will take place (no self-enrollment) Further information to registration and work hours:
VVZ CR n/a

Last Updated: 2026-02-05 15:47:20

Abstract

During this semester course, methodologies will be taught for biological-chemical enzyme evolution experiments using molecular genetic mutation technologies and in vivo selection in recombinant bacterial strains.

Objective

All technologies used for the experiments will be explained to the students 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. The 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. The course participants 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 students will present the results obtained from their individual evolution experiments at the end of the semester. We expect that during this lab course we will not only generate novel enzymes, but also gain new mechanistic insights into the investigated catalyst.

Resources

Lecture Notes

The necessary documents and protocols will be distributed to the participants during the course.

Literature

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.

General Information

Language
English
Levels
MSC
Frequency
Yearly recurring

Examination

Type
ungraded semester performance
Credit points can only be earned if the candidate (i) takes active part in the entire program, (ii) gives an oral presentation on achieved results, and (iii) turns in a final report that is accepted.Furthermore, each participant's performance will be assessed with regard to (1) effort/participation/presentation, (2) skills or acquisition of skills, and (3) quality of the report to determine whether the class is passed. Additional information will be provided at the beginning of the course.

Course Components

Type Title Time & Place Hours
practical/laboratory course Biological Chemistry A: Technologies for Directed Evolution of Enzymes
Permission from lecturers required for all students.
  • By Appointment None-None
16 h weekly

Offered In