Team:SJTU-BioX-Shanghai/A

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<li><a href="/Team:SJTU-BioX-Shanghai" class="selected">Home</a></li>
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<li><a href="/Team:SJTU-BioX-Shanghai/Project" >Overview</a><br></li>
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<li><a href="/Team:SJTU-BioX-Shanghai/Project/project1" >Membrane Protein</a><br></li>
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<li><a href="/Team:SJTU-BioX-Shanghai/Project/project2" >Accelerater</a><br></li>
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<li><a href="/Team:SJTU-BioX-Shanghai/Project/project3" >Switch</a><br></li>
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<li><a href="/Team:SJTU-BioX-Shanghai/Notebook/protocol">Notebook</a></li>
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<tr><td align="left"> <h2><b>Membrane Workshop</b></h2>
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<p>This year, SJTU-BioX-Shanghai iGEM team is trying to build a “factory” on E.coli’s membrane, where enzyme assemblies can be manipulated so that biochemical reactions can be accelerated and further controlled. </p>
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<p>We aimed at constructing a set of protein assemblies on E.coli cell membrane as carriers of various enzymes. Distinct from previous synthetic scaffold system, our device limits the reaction space to a two-dimensional surface. In such system, the membrane functions as an extensive scaffold for proteins to anchor without limitation of scaffold amount. More strikingly, we can switch direction of enzymatic reactions through external or internal signals with our device. </p>
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<p>By gathering downstream enzymes through interacting protein domains, the reaction can be accelerated sharply. Production of fatty acid was enhanced by more than 24 fold through recruiting our membrane accelerator system, which has a promising application prospect in biofuel production.</p>
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<p>Our slogan: Cluster makes it faster; interaction alters the direction</p>
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<p>In ''Membrane Rudder'' device, we successfully changed the direction of Violacein synthetic pathway through light signal. </p>
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<b>Motivation</b><br>
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<ol><li>There lacks compartment in prokaryotic cells, and thus engineered enzymes diffuse in the cytoplasm, which makes certain reactions proceed at a very low speed.  
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</li><li>Divergent biochemical pathways commonly exist in all types of organisms, but it is extremely hard to artificially control and switch the directions of these reactions.
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<b>What to do</b><br>
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<p>We aim to attach enzymes involved in certain reactions to membrane proteins in order to fulfill goals stated below:</p>
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<ol><li>To accelerate reactions in certain biochemical pathway
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</li><li>To switch the biochemical pathway from one to the other through extracellular signal control
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<h2><b>Membrane Engine (Accelerator)</b></h2>
 
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<p>As there is no compartment in prokaryotic cells, enzymes involved in a biochemical pathway diffuse all over the cytoplasm. Intermediates generated from one enzyme cannot be passed efficiently to the next due to spatial obstacles.  However, if we attach those enzymes to engineered membrane proteins which assemble together, the reactions are going to proceed much faster. </p>
 
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<ul><li>Interacting proteins fused with enzymes can decrease the distance which intermediates must travel between enzymes, improving reaction speed. 
 
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<p><i>Why do we localize the enzymes to the membrane?</i></p>
 
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<ul><li>Interaction of proteins can only be effective within a small distance. Membrane localization of the enzymes can integrate those engineered proteins to 2-dimensional scale, which would absolutely increase the possibility that potential interacting domains and ligands dimerize.
 
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<h2><b>Membrane Switch</b></h2>
 
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<p>Now that we have built a device that can speed up a biological pathway. Our next goal is to control the pathway better---- to switch the direction of certain reactions, as shown in figure 3.</p>
 
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<p>Divergent biochemical pathways commonly exist in all types of organisms, and most of those reactions are stringently and internally controlled. However, it is extremely hard to artificially control and switch the directions of these reactions. Usually there are two different products produced in divergent reactions. Sometimes we want one product, and sometimes we want the other. Using our designed device, we can change the direction by introducing different extracellular signals. </p>
 
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Latest revision as of 12:39, 18 October 2012

This year, SJTU-BioX-Shanghai iGEM team is trying to build a “factory” on E.coli’s membrane, where enzyme assemblies can be manipulated so that biochemical reactions can be accelerated and further controlled.

We aimed at constructing a set of protein assemblies on E.coli cell membrane as carriers of various enzymes. Distinct from previous synthetic scaffold system, our device limits the reaction space to a two-dimensional surface. In such system, the membrane functions as an extensive scaffold for proteins to anchor without limitation of scaffold amount. More strikingly, we can switch direction of enzymatic reactions through external or internal signals with our device.

By gathering downstream enzymes through interacting protein domains, the reaction can be accelerated sharply. Production of fatty acid was enhanced by more than 24 fold through recruiting our membrane accelerator system, which has a promising application prospect in biofuel production.

In ''Membrane Rudder'' device, we successfully changed the direction of Violacein synthetic pathway through light signal.


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