Partner 8

 Prof. Rita Bernhardt, PI

Prof. Rita Bernhardt is the Chair of Biochemistry at the Saarland University and has extended experiences in protein expression and characterization, especially of proteins involved in drug and steroid metabolism. The group has experiences in cytochrome P450 research since 1977 and in steroid hormone biosynthesis since 1988. Special attention has been given to protein-protein interactions in drug metabolizing and hormone producing systems. Her group did fundamental work in characterizing both mammalian, membrane-bound as well as bacterial, soluble steroid hydroxylase systems. Investigation of CYP106A2 is in the focus of this work since many years. Moreover, 21 cytochromes P450 of the myxobacterium Sorangium cellulosum Soce56 have been cloned, expressed and are under investigation concerning their structures and functions. In addition, improvements of P450-redox partner interactions as well as hydroxylation selectivities as prerequisite for biotechnological applications of this group of proteins have been obtained.

Institue of Biochemistry - SAAR

P4FIFTY project in the SAAR

The main aim of the work performed at SAAR will be the production of CYP106A2 mutants by site-directed mutagenesis and directed evolution with improved activities/specificities. Using the available computer model of CYP106A2, amino acids will be defined which can cause changes in the activity/ specificity of hydroxylation. Moreover, mutants will be produced using directed evolution of the protein. This way, a library of mutants of this CYP will be produced. Mutants will be screened, purified and analysed to find those with improved activity towards steroids and terpenoids and/or with changed specificity of hydroxylation. To screen the mutants, a HTS developed in our lab will be used. Moreover, CYP106A2 will be produced for crystallisation studies. The produced mutants should be a useful basis to exploit this P450 as catalysts for biotechnological processes.

Staff undertaking the work

The PhD student working in P4FIFTY at SAAR is Flòra Màrta Kiss with a Msc in Biochemical Engineering from the University of Technology and Economics Budapest. Her Master Thesis was performed in the lab of Katalin Monostory at the Chemical Research Center of the Hungarian Academy of Sciences, Laboratory of Drug-interactions.