Biochemistry of coproporphyrin ferrochelatases
- Lebensmittel, Ernährung, Gesundheit
- Biotechnologie
Abstract
Theoretical framework: Prokaryotic heme biosynthesis has been studied extensively over several decades. In 2015 this topic came back into the focus due to the discovery of the so-called “coproporphyrin-dependent” (CPD) heme biosynthesis pathway by Dailey and co-workers, which is mainly utilized by Gram-positive bacteria. The CPD pathway differs from the protoporphyrin-dependent pathway in the sequence of uroporphyrinogen to heme b transformation and the involved enzymes (UroD, CgoX, CpfC ChdC). Studies on CpfC were performed until recently using protoporphyrin IX instead of coproporphyrin III. Objectives: In-depth knowledge on the biochemistry and molecular enzymology of CpfCs is very limited. We aim at studying the protein biochemistry and elucidating the enzymatic mechanisms of the coproporphyrin ferrochelatases from (i) Firmicutes and (ii) Actinobacteria in order to understand the mechanism of catalysis, including substrate binding of both substrates and the enzymatic insertion of ferrous iron into coproporphyrin III. Methods: Biochemical and biophysical characterization of the coproporphyrin ferrochelatases (CpfCs) will be performed using multiple high-end spectroscopic methods and steady-state and pre-steady state kinetic characterization of wild-type and mutated enzymes. Further we will employ state-of the art structural biology methods to gain profound knowledge of the enzyme’s structure to the very detail, such as protonation states (by neutron crystallography). Innovation: The CPD heme biosynthesis pathway was recently described in literature and many questions about the enzymes’ mechanisms and interactions are unanswered. It is essential for Gram-positive bacteria and a few intermediate and Gram-negative bacteria. Understanding the biochemistry of the involved enzymes, focussing on CpfCs, and the overall regulation of heme biosynthesis, uptake and degradation of these bacteria is a highly important basic research question. This project will provide a strong basis for the future development of new therapeutics against pathogenic Gram-positive bacteria.
Project staff
Stefan Hofbauer
Ass.Prof. Priv.-Doz. Dipl.-Ing. Stefan Hofbauer Ph.D.
stefan.hofbauer@boku.ac.at
Tel: +43 1 47654-77258
Project Leader
01.07.2020 - 31.01.2025
Paul Georg Furtmüller
ao.Univ.Prof. Dipl.-Ing.Dr.nat.techn. Paul Georg Furtmüller
paul.furtmueller@boku.ac.at
Tel: +43 1 47654-77277
Project Staff
01.07.2020 - 31.01.2025
Thomas Gabler
Dipl.-Ing. Thomas Gabler BSc. PhD.
thomas.gabler@boku.ac.at
Tel: +43 1 47654-77278
Project Staff
01.07.2020 - 31.01.2025
BOKU partners
