Team:Bielefeld-CeBiTec/Results/CO2-fixation

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The particular aim of the second module is to realize the carbon dioxide fiaxtion in <i>E. coli</i>. For this approach all items, like the <a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Results/CO2-fixation/Calvin-Cycle" target="_blank">Sedoheptulose-1,7-Bisphpospahtase</a>, the <a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Results/CO2-fixation/RuBisCO" target="_blank">Ribulose 1,5-bisphosphate carboxylase/oxygenase</a>  (RuBisCO) and the mechanism of <a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Results/CO2-fixation/RuBisCO" target="_blank">carbon dioxide fixation</a> a tested separetly in various approaches. For the optimization of the carbon dioxide fixation under aerobic growth conditions we investigate the anerobic microcompartment from <i>Halothiobacillus neapolitnaus</i>, called the <a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Results/CO2-fixation/Carboxysome" target="_blank">carboxysom</a>.<br>
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The particular aim of the second module is to implement the carbon dioxide fiaxtion in <i>E.&nbsp;coli</i>. Therefore we selected the Calvin cycle (figure 1) and used a bottom up approach. All heterologous expressed components, like the <a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Results/CO2-fixation/Calvin-Cycle" target="_blank">sedoheptulose-1,7-bisphosphatase (<i>glpX</i>)</a>, the phosphoribulokinase (<i>prkA</i>) , the <a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Results/CO2-fixation/RuBisCO" target="_blank">ribulose-1,5-bisphosphate carboxylase/oxygenase</a>  (RuBisCO) were tested separately in various experiments. The RubisCO is known to function best under high CO<sub>2</sub> concentration. To accomplish optimal conditions for the RubisCO in a very local enviroment a microcompartiment from <i>Halothiobacillus&nbsp;neapolitanus</i>, which is called carboxysome, was constructed in <i>E.&nbsp;coli</i>.
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<font size="2" style="text-align:center;"><b>Figure1:</b> Schematic representation of the calvin cylce. The reaction shwon in green can be realized by enzymes that naturally exist in <i>E. coli</i>, while the red one need to be expressed heterologous to fullfill the whole calvin cycle in <i>E. coli</i>.</font>
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<font size="2" style="text-align:center;"><b>Figure 1:</b> Schematic representation of the Calvin cylce. The reactions shown in green can be catalyzed by enzymes that naturally exist in <i>E.&nbsp;coli</i>, while the red ones need to be expressed heterologous to enable the whole <a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Results/CO2-fixation/Calvin-Cycle">Calvin cycle</a> in <i>E.&nbsp;coli</i>.</font>
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The different results for all three enzymes are mentioned in the <a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Results/CO2-fixation/Calvin-Cycle" target="_blank">Calvin-cycle</a> section. One important step for the carbon dioxide fixation is the <a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Results/CO2-fixation/RuBisCO" target="_blank">RuBisCO</a> (Ribulose 1,5-bisphosphate carboxylase/oxygenase). We decided to transform DNA sequences into <i>E. coli</i> which encode the <a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Results/CO2-fixation/Carboxysome" target="_blank">carboxysome</a>. Due to its special properties this microcompartiment is very usefull for the carbon dioxide fixation.
 
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Latest revision as of 02:57, 18 October 2014


Module II - Carbon Dioxide (CO2) Fixation

The particular aim of the second module is to implement the carbon dioxide fiaxtion in E. coli. Therefore we selected the Calvin cycle (figure 1) and used a bottom up approach. All heterologous expressed components, like the sedoheptulose-1,7-bisphosphatase (glpX), the phosphoribulokinase (prkA) , the ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBisCO) were tested separately in various experiments. The RubisCO is known to function best under high CO2 concentration. To accomplish optimal conditions for the RubisCO in a very local enviroment a microcompartiment from Halothiobacillus neapolitanus, which is called carboxysome, was constructed in E. coli.



Figure 1: Schematic representation of the Calvin cylce. The reactions shown in green can be catalyzed by enzymes that naturally exist in E. coli, while the red ones need to be expressed heterologous to enable the whole Calvin cycle in E. coli.

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