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Team:Bielefeld-CeBiTec/Results/rMFC
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| - | Our first module deals with the construction of an <i>E. coli</i> strain, which is able to accept electrons stimulating its metabolism. We focused on the indirect electron transfer via <a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Project/rMFC/Mediators">mediators</a>, which are reduced at the electrode in a <a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Project/rMFC/ReactorSystem#DesignReactorSystem">electrobiochemical reactor</a> and then reoxidized again inside bacterial cells. | + | Our first module deals with the construction of an <i>E. coli</i> strain, which is able to accept electrons stimulating its metabolism. We focused on the indirect electron transfer via <a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Project/rMFC/Mediators">mediators</a>, which are reduced at the electrode in a <a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Project/rMFC/ReactorSystem#DesignReactorSystem">electrobiochemical reactor</a> and then reoxidized again inside bacterial cells. The electron transfer system consists of different steps. First the reduced mediator has to cross the outer membrane of the <i>E. coli</i> cell. For that we are going to use the outer membrane porine OprF (<a href="http://parts.igem.org/wiki/index.php/Part:BBa_K1172507">BBa_K1172507</a>) provided by <a href="https://2013.igem.org/Team:Bielefeld-Germany/Project/Porins">iGEM Team Bielefeld-Germany 2013</a>. |
| + | Crossing the periplasmatic space, the mediator adsorbs at the inner membrane of the <i>E. coli</i> cell. Electrons have to be transferred into the cytoplasm, but the mediator should not enter the cell, because regeneration at the electrode is absolutely necessary. <br> | ||
| + | We are going to modify the metabolic pathway of fumarate by a knockout of the C4 carboxylate antiporter DcuB in <i>E. coli</i> and overexpression of different fumarate reductases. Furthermore the outer membrane porine OprF had to be integrated into the bacterial chromosome to ensure a constitutive expression of OprF and reduce the plasmid overload of bacterial cells. | ||
Revision as of 22:34, 17 October 2014
Module I - Reverse Microbial Fuel Cell (rMFC)
Our first module deals with the construction of an E. coli strain, which is able to accept electrons stimulating its metabolism. We focused on the indirect electron transfer via mediators, which are reduced at the electrode in a electrobiochemical reactor and then reoxidized again inside bacterial cells. The electron transfer system consists of different steps. First the reduced mediator has to cross the outer membrane of the E. coli cell. For that we are going to use the outer membrane porine OprF (BBa_K1172507) provided by iGEM Team Bielefeld-Germany 2013.
Crossing the periplasmatic space, the mediator adsorbs at the inner membrane of the E. coli cell. Electrons have to be transferred into the cytoplasm, but the mediator should not enter the cell, because regeneration at the electrode is absolutely necessary.
We are going to modify the metabolic pathway of fumarate by a knockout of the C4 carboxylate antiporter DcuB in E. coli and overexpression of different fumarate reductases. Furthermore the outer membrane porine OprF had to be integrated into the bacterial chromosome to ensure a constitutive expression of OprF and reduce the plasmid overload of bacterial cells.
