Grit Daniela Straganz
Johannes Niederhauser: Nonheme-Iron Dioxygenases: From Structure-Function Relationships via Redesign towards Whole-Cell-Factories
Sarah Maria Pratter: Expression and Engineering of Oxygenases with New Catalytic Properties
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Grit Daniela Straganz
Institute of Biotechnology and Biochemical Engineering
Technische Universität Graz
phone: +43 316 873 8414
fax: +43 316 873 8434
Exploring and Expanding the Catalytic Potential of a Non-heme Fe(II) Dependent Dioxygenase
O2 metabolism is crucial for almost all terrestrial life. Nature evolved enzymatic structures to allow for the controlled reaction of O2 with organic substrates. Enzymatic nonheme metal- and particularly nonheme-Fe(II) centres have raised much interest in recent years, because of the unparalleled diversity of reactions they catalyze. G. Straganz’s research focuses on the reactivity of these enzymatic metal-centres and the role of both the Fe(II) centre and the protein environment (i) in the mechanism of dioxygen activation and (ii) in the fate of the subsequent reactive oxygen species, which causes the diversification of reactions. A set of biochemical, biophysical and molecular genetic methods is used in order to elucidate the reaction mechanisms at the metal-centre and to characterize the role of the protein environment. This knowledge forms the basis for the engineering of enzymes with novel catalytic properties.
Laboratory know-how and infrastructureThis workgroup’s strength is in the biochemical and kinetic characterization of dioxygen dependent metalloenzymes, particularly of nonheme Fe(II) dependent dioxygenases, whereby also anaerobic conditions may be employed. An HPLC is also available, which has an UV and an RI detector, for the routine characterization of the reaction products. Site-directed mutagenesis, heterologous overexpression and purification of proteins from E.coli are routine procedures. The Institute of Biotechnoloy and Biochemical Engineering is well equipped for all these tasks. It has 5 spectrophotometers, a fluorimeter, a stopped flow instrument that can act as a spectrophotometer and a fluorimeter, a glove box for working anaerobically and a Clarke electrode or monitoring O2 dependent reactions. Shakers, a French press, Ultracentrifuge, Äkta-System and gel-electrophoresis for the preparation of protein are also available. The institute has furthermore rooms dedicated to genetic work, with PCR machines and a DNA-Gel-Electrophoresis dark room. A computer for in silico docking analyses is available. Autodock 4, a widely used free software package (Scripps Insititute, La Jolla) that allows induced-fit-docking is already employed by the applicant and group members.