Team:Cologne-Duesseldorf/Test4

  • 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.