OBJECTIVES
OBJECTIVE 1.
Rational design driven site-directed mutagenesis for identification of D-selectivity modulating residues:
Related work-packages:
- WP1. Site-directed mutagenesis driven by rational design concepts
- WP2. Ala-scan of active site residues combined with monodental substrate fixation
- WP3. D-selectivity measurements
OBJECTIVE 2.
Iterative saturation mutagenesis – CASTing at D-selectivity modulating sites:
Related work-packages:
- WP1. Saturation mutagenesis
- WP2. Activity screens of transformants from saturation mutagenesis
OBJECTIVE 3.
Functional and structural characterization of novel PALs of reversed D-selectivity:
Related work-packages:
- WP1. Expression, isolation and purification of the PALs of D-activity
- WP2. Functional characterization of PALs of enhanced D-selectivity
- WP3. Structural characterization of the novel recombinant PALs of enhanced D-selectivity
OBJECTIVE 4.
Development of PAL-based biotechnologies for valuable D-phenylalalanines:
Related work-packages:
- WP1. Reaction engineering using D-PALs as whole cell biocatalysts
- WP2. Engineering of biotransformations with purified, immobilized D-PALs