Team:Cologne-Duesseldorf/jan

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Secretion

Downstream processing is not only time consuming but also cost and energy intensive. Therefore, we aim to simplify the purification of compounds produced in our artificial compartment. We used a concept based on the peroxicretion described by Sagt and colleagues <a href=" https://www.ncbi.nlm.nih.gov/pubmed/19457257"> (Sagt et al, 2009) </a>.

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Introduction

Downstream processing is a very important part of industrial biological compound production. For most biotechnological produced compounds, it is the most expensive part of the production <a href=" https://www.ncbi.nlm.nih.gov/pubmed/11602307 "> (Keller et al, 2001) </a> . One step to decrease the costs is to secrete the products into the supernatant <a href=" https://www.ncbi.nlm.nih.gov/pubmed/23385853"> (Berlec et al, 2013) </a>. After secretion, it is possible to remove most cellular compounds from valuable products with one simple centrifugation step. Due to this, secretion is not only a great tool for a compartment toolbox but also has an economic value.
In regards to the whole project, this is an important part for making the compartment more applicable. Through it we go a step further by thinking about the extraction of products after production.
At the end of this sub project it should be possible to secrete every compound produced in the modified compartment to the supernatant. This is not trivial because peroxisomes, which are the basis of our compartment do not possess a known natural secretion mechanism.
We overcome this problem by using the "peroxicretion" concept of Sagt and colleagues <a href=" https://www.ncbi.nlm.nih.gov/pubmed/19457257"> (Sagt et al, 2009) </a>. They used a v-SNARE (vesicle- synaptosome-associated-Soluble N-ethylmaleimide-sensitive-factor Attachment REceptorprotein) fused to a peroxisomal membrane-protein to secrete the content of peroxisomes. V-SNAREs interact with the t-SNARE (target synaptosome-associated-SNARE) at the cell membrane, which leads to an fusion of the vesicle with the membrane <a href=" https://www.nature.com/nrm/journal/v2/n2/full/nrm0201_098a.html"> (Chen et al, 2001) </a>.