Enzyme/protein engineering: CELL BIOLOGY EDUCATED PROTEIN DESIGN AND TESTING

DESIGN strategies to produce high levels of recombinant proteins and peptides in plants as bioreactors according to an in silico pipeline

The known structural features of the protein of interest (POI) are used for in silico modelling of the POI, and the choices of vector, target sequence, codon-usage, expression system and purification strategy are tailored to the required POI properties (e.g. stability, purity, or alleviation of allergenic/toxic features).

TESTING the different strategies by transient expression in plant systems

In vivo test of the engineered POI using rapid, reliable transient expression systems (plant cell protoplasts, leaf agroinfiltration, yeast or bacterial expression), depending on the POI features. POI stability (half-life) and subcellular localization as well as co- and post-translational modifications are monitored by microscopy and biochemical/biophysical analysis.

Optimisation of recombinant POI(s) sequence(s), expression system and expression strategy is carried out by testing for POI yield, its intracellular localization/stability and monitoring host physiology and/or transcriptomics of the expression host during POI expression (for plants, this is performed in different tissues).

The best protein engineering strategy will be used for stable production of POI in plants.

Vectors for transient and permanent expression containing the POI of interest are provided to the customers as the final product. For details see https://www.youtube.com/watch?v=Lj3v6G6YTY4