Structure-based engineering of aspartate ammonia lyase

Project description

It concerns a master research project aimed at the engineering of aspartate ammonia lyase (aspartase) variants that can be used as biocatalysts for the synthesis of various pharmaceutically important beta-amino acids. This protein engineering attempt will be guided by the recently solved crystal structure of aspartase in complex with its natural substrate (http://www.ncbi.nlm.nih.gov/pubmed/21661762). The major benefit of such a structure-based evolution strategy lies in the reduction of the sequence space that has to be covered in order to identify valuable mutant variants. The student will start with the construction of a large library of aspartase variants, using saturation mutagenesis at several combined positions in the substrate-binding pocket. Screening or selection will be used to obtain active aspartase variants, of which the biocatalytic properties will be investigated. The scope of beta-amino acids that can be synthesized using the newly designed aspartases will be explored by using UV spectroscopy, HPLC-MS, and NMR.

Organization

The master research project will be conducted in the Pharmaceutical Biotechnology group of Prof.dr. G.J. Poelarends. Our laboratory is well equipped for molecular biology, enzymology, protein engineering, and biocatalysis work. We offer a stimulating research environment for a student interested in these topics. There are also good contacts with the pharmaceutical industry.

Requirements

We are looking for highly motivated students interested in performing state-of-the-art protein engineering work.

Contact

For further information you can contact Prof.dr. G.J. Poelarends, Dept. of Pharmaceutical Biology, GRIP, building 3215-room 9.21, Antonius Deusinglaan 1, 9713 AV GRONINGEN.