We were able to design and successfully test an orthogonal peroxisomal protein import mechanism for the peroxisome in S. cerevisiae.
By decorating the peroxisomes with the v-SNARE Snc1 we successfully secreted their entire contents
With two different sensors we were able to efficiently measure the pH and the redox potential inside our yeast peroxisomes.
Via fluorescence microscopy we verified that the integration of new membrane proteins into the peroxisomal membrane is possible.
By successfully translocating the required enzymes for the metabolic pathways of nootkatone and violacein into the peroxisome and actually synthesizing the latter, we developed a proof of concept for our toolbox
We successfully implemented a way of customizing the size and number of the peroxisomes into our toolbox.
With a high throughput assay we characterized the import efficiency of different PTS2 sequences.
To get a better understanding of possible problems and pitfalls of our metabolic engineering concepts we extensively modeled the whole nootkatone pathway and the benefits of it being translocated inside our compartment.
For our planned optogenetic experiments we designed an affordable lightbox which can easily be assembled in a short time.
