RAtional REdesign of Phenylalanine Ammonia-Lyases for reversing their natural selectivity (RARE-PAL)


The aim of the RARE-PAL project is the reversal of the natural L-enantioselectivity of phenylalanine ammonia-lyases (PALs) by the aid of protein engineering. The five-membered project team intends to perform a highly interdisciplinary research, combining molecular biology, enzymology, biocatalysis, biochemistry, biochemical process engineering, resulting in advanced biotechnologies for the production of D-phenylalanine analogues of high synthetic value.

The project team proposes an original rational design concept based on our previous results, which combined with the employment of last generation protein engineering techniques provides the basis for the reversal of PALs enantioselectivity.

Through its activities the project:

  • i) will identify the D-selectivity modulating residues of PcPAL using the novel rational design concept
  • ii) will apply the emerged protein engineering techniques, such as iterative saturation mutagenesis combined with high-throughput D-PAL-activity screens, to develop PALs with D-selectivity
  • iii) develops expression systems and isolation procedures of the novel PALs of reversed selectivity
  • iv) employs the developed D-PALs, as whole-cell or immobilized biocatalysts, to provide batch and continuous flow biotechnologies for the production of D-phenylalanines of high industrial interest