Difference between revisions of "Team:Aix-Marseille/Design"

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    <h2 class="title wow bounce in up"><span style="color:#8E3B8C"><span style="font-family:Armalite Rifle">Purification</h2></span></span>
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    <p class="space100"><div align="justify"><span style="font-family: Courier New">We added a His-tag following the sequence of genes coding our proteins.
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Using columns containing nickel ions, His-tagged proteins are bound with high specificity and affinity. Other proteins pass through the matrix during the washing step. His-tagged proteins are eluted using imidazole buffer.
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Revision as of 10:56, 12 September 2015

Chew fight

Project design

Our project this year is to work on the chewing gum degradation. Our research on this topic reveals that chewing gum is composed of different compounds. They are hydrophilic or hydrophobic. When you chew a chewing gum, the hydrophilic part is solubilized by your saliva. Once done, you throw away the hydrophobic part called the gum base. The gum base is composed of polymers : polyisoprene and butadiene.

So we searched for enzymes which are able to degrade polymers. We found two enzymes: lipoxygenases and laccases.

At the beginning, our main problem was that laccase is able to oxidize the ruthenium. But we did not want to use it in our project because of its scarcity, its toxicity and impact on the environment. We found that the cytochrome C which uses iron could replace it and be oxidized by laccases. We decided to try and asked if it could help to degrade our polymers. We selected several enzymes that we believed could be great to use in E.coli strain.

Our strategy

Our strategy is to use a reduced laccase which will be oxidized by the oxygen present in the air.

To return at a reduced state, laccase will oxidize the cytochrom C previously excited by light. A superoxide anion radical will be formed.

When the cytochrome C will come back to its reduced state and the superoxide anion radical to oxygen, the polymers will be in a radical state.

Thanks to oxygen, it will form an epoxide that will be hydrolyzed and become a diol.

So we think that the polymer degradation will be more efficient by this method.