Membrane Integration
introducing new proteins to the membrane
Introduction
Compartmentation has been one of nature’s most effective tools for more than a billion years. The tremendous versatility of organisms we see today is only possible because cells have developed the ability of translocating various metabolic processes to subcellular compartments, thereby sequestering them from others. Our project is about harnessing the full potential of this awesome mechanism. What used to have to evolve over millions of years can now be directly controlled and customized throug use of our toolbox. Towards this aim we worked on many different sub projects, each targeting a different aspect of compartment customization. Below you will find a description on all of them.
Design and modeling
We have chosen yeast peroxisomes as our chassis for designing synthetic organelles. They are very resistant, have a modifiable import mechanism and are expendable under optimal conditions. We will customize the import machinery of peroxisomes in yeasts in order to regulate the biomolecule import into these compartments. To do so, we modify the TPR-region of the peroxisomal target protein receptor PEX5 with modeling, so that it only recognizes a single new designed peroxisomal import signal. The most promising modified PEX5s will be implemented into the actual peroxisome.
Real world application
As a proof of concept for our compartimentation strategy we intend to establish the Nootkatone pathway inside the peroxisome. Nootkatone is a natural compound found inside the peel of the grapefruit, which gives it its characteristic taste and smell. In addition, Nootkatone is a natural repellent for mosquitoes and ticks that is already being commercially used and industrially manufactured. Unfortunately, the production costs are extremely high, because it has to either be extracted from the peels of millions of grapefruits or synthesized inside of yeast. The problem is that the Nootkatone pathway is toxic for yeast and the efficiency is rather low. Here our compartmentation comes into play: we plan to implement the whole pathway into the modified peroxisome to prove, that we have transformed a peroxisome to an independent compartment with all the features required by us
