Partner 3

Institute of Technical Biochemistry, Universitaet Stuttgart

The Institute of Technical Biochemistry is led by Prof. Bernhard Hauer and applies engineered enzymes to biocatalysis focusing on oxidoreductases, inter alia P450 monooxygenases, with the target of providing industry with working solutions. The laboratories are equipped to implement analytical chemistry, microbiology, protein biochemistry and molecular biology. Research is focused on cloning, expression, characterization and optimization of enzymes for industrial applications. The bioinformatics group of Prof. Jürgen Pleiss focuses on investigating the molecular basis of enzyme properties and to design improved biocatalysts. Two complementary approaches are combined, the systematic analysis of sequences and structures of large protein families by establishing family-specific databases, and the molecular modeling of proteins and protein complexes using molecular dynamics simulations.

Prof. Dr. Bernhard Hauer, PI

Bernhard Hauer is the head of the Institute of Technical Biochemistry. After many years working in the chemical industry BASF SE (Germany), in 2009 he became professor at the Universitaet Stuttgart. His research interests cover the provision of novel biocatalysts to further expand the spectrum of accessible biocatalytic reactions along with the development of methodologies to create novel materials.

Prof. Dr. Jürgen Pleiss, PI

Jürgen Pleiss has a long experience in the design of P450 mutants with improved selectivity. Recently, a highly focused, minimal mutant library of bacterial P450 BM-3 was established and successfully screened for variants with improved selectivity. By combining protein modeling and a systematic analysis of the in-house developed Cytochrome P450 Engineering Database (CYPED), promising candidates are further improved by enzyme design for activity, selectivity, and stability. The CYPED will also be applied in screening P450 gene diversity in collaboration with other partners.

P4FIFTY project

The work performed at USTUTT within P4FIFTY will focus on the development of methods for using cytochrome P450 monooxygenases (P450) for terminal hydroxylation reactions of fatty acids and alkanes on an industrial scale. The chosen targets are key chemical building blocks with numerous industrial applications; however, activity and selectivity of these enzymes towards alkanes and fatty acids of industrial interest are often quite low. To overcome this limitation, USTUTT will aim for the development and screening of P450 mutant libraries for enzymes with improved activity and selectivity. In addition, USTUTT will investigate a combination of P450 mutant enzymes and cofactor recycling systems in highly efficient whole cell biotransformation systems. Further work include the implementation of enzyme diversity studies with partner institutions and the application of molecular modeling approaches combined with rational protein design methods using the in-house developed “Cytochrome P450 Engineering Database” (CYPED).

Staff undertaking the work

Sandra Notonier is working in P4FIFTY at USTUTT. Sandra obtained her MSc in biotechnology from the Ecole Superieure d´Ingenieurs de Luminy, France. Prior to her PhD she has completed internships in metabolic engineering at LS9 Inc. (USA) and in the construction of biosensors at the French National Centre for Scientific Research CNRS (France).

The PhD thesis of Lukasz Gricman in P4FIFTY focuses on establishing a modeling strategy for P450 enzymes. Lukasz Gricman will apply molecular modeling to design P450 enzyme variants with improved stability, activity, and selectivity, and the CYPED for data mining and in silico screening for new P450 enzymes.