Prof. Dr. Ulrich Schwaneberg
RWTH Aachen
Lehrstuhl für Biotechnologie
Worringerweg 3
52056 Aachen
Lehrstuhl für Biotechnologie
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Master Thesis
Identification, characterization and activity tests of ketsteroid5β-reductase.
Introduction:
The goal of the project would be identification of most stable, active and substrate specific
ketosteroid-5β-reductase. Ketosteroid-5β-reeducates are involved in steroids metabolism and were
found in many eukaryotic organisms like human, rat, yeast, mouse etc. Several proteins from this
family were successfully overexpressed in E.coli. X-ray structure of human enzyme is already
known. Eukaryotic ketosteroid-5β-reductases accept broad range of substrates, but kinetic
properties of this enzyme have not been thoroughly studied.
Description of project:
The project is done in collaboration with industrial partner - PharmaZell GmbH.
Following work should be done in frame of master work - 6 months. The student should have
background in molecular biology and biochemistry.
Working package 1: Selection of several eukaryotic genes of ketosteroid-5β-reeducates (wt or
mutants), which can be successfully overexpressed in bacterial hosts.
Working package 2: Cloning of selected genes and overexpression of enzyme in bacterial
expression system (E.coli, Gram positive strains). Stability tests of the enzyme must be performed.
Working package 3: Verification of activity and substrate specificity of the 5β-reductase. Different
substrates delivered by PharmaZell should be tested with whole cells and cell extracts (optionally
with purified enzymes). Optimization of catalytic conditions should be performed. Analytical part will
be performed by PharmaZell.
Selected publications:
1.
2.
3.
Crystal Structure of Human Liver Δ4-3-Ketosteroid 5β-Reductase (AKR1D1) and Implications for Substrate
Binding and Catalysis
Luigi Di Costanzo, Jason E. Drury, Trevor M. Penning and David W. Christianson. J Biol Chem. 2008 June 13;
283(24): 16830–16839.
Substrate specificity and inhibitor analyses of human steroid 5β-reductase (AKR1D1) Mo Chen, Jason E. Drury,
Trevor M. Penning, Steroids, Vol 76, April 2011, Pages 484–490
Contact:
Dr. Anna Joelle Ruff, Lehrstuhl für Biotechnologie, Worringerweg 3, 52056 Aachen, Raum 4.133,
Tel.: 0241 80 23604, aj.ruff@biotec.rwth-aachen.de