Prof. Christof Holliger, Dr. Julien Maillard, Mathilde Willemin, N.N.
Swiss National Science Foundation & EPFL
January 2017 –December 2021
PD Dr. Antonio Pierik, University of Marburg, Germany; Dr. T. Drepper, University of Düsseldorf, Germany; Prof. Ines A. Pereira, Universidade Nova de Lisboa, Oeiras, Portugal
Organohalide respiration (OHR) is an energy metabolism allowing certain bacteria to use organohalides as terminal electron acceptor. The biochemistry of the OHR metabolism still remains relatively elusive since only a few reductive dehalogenases (RDases), the key catalytic enzymes in OHR, have been studied. The biochemical characterization of other gene products encoded in reductive dehalogenase (rdh) gene clusters remain rather unexplored. Among these, two enzyme families, RdhC and RdhK, are the main targets we would like to investigate with this proposal.
We will apply state-of-the-art biochemistry methods to study gene products of rdh gene clusters other than RdhA and show their involvement in the OHR metabolism. PceC will be characterized by using the recently in-house produced FMN-binding domain of PceC (PceCFD) and by producing the whole protein in a bacterial host that contains an intracytoplasmic membrane system allowing the production of membrane proteins such as PceC. The RdhK proteins will be studied by producing hybrid proteins harboring the N-terminal organohalide-binding domain of new RdhK proteins fused to the C-terminal DNA-binding domain of a characterized RdhK protein. Once the organohalide partners are identified, surface plasmon resonance (SPR) spectroscopy will be used to screen for DNA targets of selected RdhK proteins.