Protein engineering of sortases

The aim of our research is:

• To use sortases as a target for the development of new antibiotic strategies

• To engineer sortases to catalyse the transpeptidation reaction in vitro

Sortases are housekeeping enzymes displayed on the surface of Gram-positive bacteria, including Staphylococcus aureus – a leading cause of health care concerns. As housekeeping enzymes sortases perform essential metabolic functions for the purpose of survival. On the cell surface, sortases are involved in ‘pasting’ virulence factors on the peptidoglycan of the bacteria in a transpeptidation reaction, making them essential for pathogenesis. Therefore, sortases make an interesting target for the development of new antibiotic strategies that would help in the fight against antibiotic resistance.

On the other hand, the transpeptidation reaction sortases catalyse can be used in vitro to stick for example toxic molecules onto a protein. Sortases recognise small peptide motifs; by first breaking and then making new peptide bonds between these motifs sortases can create new molecules or molecular formats that did not exist before and that cannot be produced by nature. Therefore, sortase-mediated transpeptidation has great potential to be used in a variety of biotechnology-based applications. We are working on the optimisation of the enzyme by applying mutagenesis

In our laboratory we work on the molecular (DNA, RNA) and macromolecular (protein) level. We use different laboratory techniques starting from the fundamental molecular biology (PCR, cloning, transformation), microbiology and biochemistry methods (selection, protein purification) up to novel genome-editing and high-throughput screening techniques.

For more information please contact: Magdalena Wojcik, m.wojcik rug.nl ;

phone number: 050 3633039