Team:York/Project

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<h1>The Challenge</h1>
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<h2>Our project</h2>
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<p>This year, iGEM York are working on a bioremediation project, involving the decontamination of wastewater. Our aim is to increase sulphate uptake in E.coli and chelate Cadmium (a toxic heavy metal) through the use of metal-binding proteins called phytochelatins.  
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<p><h2>To remove Cadmium from water</h2></p>  
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Our project is divided into two main approaches that are interconnected: </p><ul>
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<li><p>The increase of sulphate uptake (using an exogenous sulphate transporter from Bacillus) and its conversion into cysteine by tweaking the cysteine synthesis pathway.</p></li>
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<p>Both cadmium and sulfate compounds can be found in wastewater and is produced from processes such as electroplating. If these contaminants are not removed from the environment, they can have a detrimental impact upon living organisms. For example, if cadmium is consumed by mammals over a long period of time, it can cause health problems such as Itai-Itai disease (cadmium poisoning). This year, our project at iGEM York is focusing on increasing the uptake of cadmium and sulfate in our chosen chassis <i>E. coli</i>. The project has two main, interlinked approaches:</p>
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<li><p>The increase of cadmium ion uptake (upregulation of cadmium transporters) and chelation (stabilisation of the metal by metal binding proteins). </p></li></ul><br>
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<p>The metal binding proteins are the link between the two approaches as they contain sulphur rich cysteine residues which will act as sinks for the cysteine overproduction.
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<li><p>Firstly, the increased uptake of sulfur using an exogenous sulfate transporter from <i>Bacillus</i>.</p></li>
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The metal binding proteins are divided into two main categories: </p><ol>
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<li><p>Secondly the increased uptake and chelation of cadmium ions through the use of metal-binding proteins, to produce a potentially recoverable metal product.</p></li>
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<li><p>Phytochelatins (synthesized by a phytochelatin synthetase in a stepwise reaction – so we are working with an exogenous Phytochelatin synthetase gene in E coli)</p></li>
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<li><p>Synthetic phytochelatins (engineered and directly translated from the DNA/RNA sequence without the need of a synthase).</p></li></ol><br>
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<p>Our system is regulated by the metal concentration in the environment. If the concentration reaches the threshold of our cadmium inducible promoter (pYoda) sensitivity then it will activate the whole system. Thus, our system prevents the overproduction of cysteine when Cadmium is not found at high concentrations.</p>
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<p><h2>How are these two processes intertwined?</h2></p>
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<h2>How would our project be used in practical applications?</h2>
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<p>In the future, we could imagine our bacteria inside of a semipermeable membrane in a river or water treatment plant with contaminated water flowing through. The high metal concentration would activate the system and our bacteria would start harvesting the pollutants. It could be coupled to a biosensor and then, when a Cadmium concentration threshold is reached the bacteria could change colour, from white to red. This colour change would indicate that it is time for “harvesting”. When this colour change is elicited, the cells could either be lysed by a chemical process or even induced to do it by a kill switch and the metal could then recovered and sold for commercial gain making decontaminating water profitable!<br><br>
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In the future, we wish to investigate chelating other metals that are also found in waste water that have greater monetary value. Vanadium for example, could be chelated in a similar process to our system outlined above and sold on.</p>
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<p>We are tweaking the cysteine biosynthesis pathway, allowing the over-expression of cysteine and the utilisation of the sulfate that accumulates inside our cell. In addition, we are tweaking another system, the production of cysteine-rich phytochelatins. We are over-producing these metal-binding proteins, and thus creating a sink for the cysteine that is produced by our cell. These metal-binding proteins are also responsible for chelating the cadmium that our cell takes up from the environment.</p>
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<p><h3>How is our system regulated?</h3></p>
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<p>We have designed our system to be regulated by the concentration of cadmium in the environment. If the concentration of cadmium surpasses the sensitivity threshold of pYoda (cadmium-inducible promoter)then it will activate our system and as a result, our genes will be expressed. Our system prevents the overproduction of cysteine when cadmium is at low concentrations.</p>
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<!-- <img src=https://static.igem.org/mediawiki/2014/4/4f/York_Cysteine_2.png>
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<p><b>Structure of L-Cysteine</b></p> -->
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<img src="https://static.igem.org/mediawiki/2014/9/99/L-cysteine.png" class="img-responsive" style="width:350px;">
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<p><b> Synthesis of L-Cysteine</b></p></div></div>
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<a href="https://2014.igem.org/Team:York/Article">
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<h3 class="text-center"><i>EcoCADMUS</i>: The grisly story behind the name.</h3></a><br>
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Latest revision as of 03:33, 18 October 2014

Team York 2014


The Challenge

To remove Cadmium from water

Both cadmium and sulfate compounds can be found in wastewater and is produced from processes such as electroplating. If these contaminants are not removed from the environment, they can have a detrimental impact upon living organisms. For example, if cadmium is consumed by mammals over a long period of time, it can cause health problems such as Itai-Itai disease (cadmium poisoning). This year, our project at iGEM York is focusing on increasing the uptake of cadmium and sulfate in our chosen chassis E. coli. The project has two main, interlinked approaches:

  • Firstly, the increased uptake of sulfur using an exogenous sulfate transporter from Bacillus.

  • Secondly the increased uptake and chelation of cadmium ions through the use of metal-binding proteins, to produce a potentially recoverable metal product.

How are these two processes intertwined?

We are tweaking the cysteine biosynthesis pathway, allowing the over-expression of cysteine and the utilisation of the sulfate that accumulates inside our cell. In addition, we are tweaking another system, the production of cysteine-rich phytochelatins. We are over-producing these metal-binding proteins, and thus creating a sink for the cysteine that is produced by our cell. These metal-binding proteins are also responsible for chelating the cadmium that our cell takes up from the environment.

How is our system regulated?

We have designed our system to be regulated by the concentration of cadmium in the environment. If the concentration of cadmium surpasses the sensitivity threshold of pYoda (cadmium-inducible promoter)then it will activate our system and as a result, our genes will be expressed. Our system prevents the overproduction of cysteine when cadmium is at low concentrations.

Synthesis of L-Cysteine

EcoCADMUS: The grisly story behind the name.


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