Team:Bielefeld-CeBiTec/Results/CO2-fixation

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The second module aims to change <i>E. coli</i> in a way that it binds carbon dioxide. This changes this bacterium from heterotroph to autotroph. To achieve this we want to establish the Calvin-cycle. <i>E. coli</i> has all enzymes for the Calvin-cycle except of three.<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="1" style="text-align:center;"><b>Figure1:</b> Missing enzymes in Calvin cycle</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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<a href="https://2014.igem.org/Team:Bielefeld-CeBiTec/Notebook/Journal/CO2-fixation">Here</a> you will find information about the execution of our experiments.
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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.