Research Internship
Marburg An Der Lahn, Hesse, Germany
Research Project at the MPI working on improving the CETCH cycle, a synthetic pathway for carbon dioxide fixation developed by Prof. Dr. T. Erb and his group. During my internship I focused on engineering specific mutations to a Succinyl-CoA reductase enzyme (SucD), to increase specificity, i.e. reduce the side-reactivity with other CoA-esters that are present as CETCH cycle intermediates. We worked with SucD variants from both Clostridium kluyverii and Clostridium difficile, where we identified amino acid residues in and around the active site, that could be mutated to improve specificity while aiming to retain activity. The work was published in ACS Biochemistry (see publications).Skills acquired during the work:Synthesis of different CoA esters.Rational design and site directed mutagenesis by quick change PCR.His-tag purification of the mutants and the wildtypes.Simple “Kinetics“ of the mutants and the wildtypes with the purified CoA esters to determine enzyme activity.Complete kinetics and crystallization of successful, active mutants.