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<h5> Our Design </h5> | <h5> Our Design </h5> | ||
− | <p>For our project, we have designed a plasmid that secretes MHETase and the double mutant PETase to increase the rate at which PET is degraded compared to the previous PETase sequence. We inserted the plasmid into an E. coli MG1655 strain. Because of the toxicity of ethylene glycol, a second plasmid was designed to allow the bacteria to break down the ethylene glycol and utilize its products as a carbon source. These enzymes include glycolaldehyde reductase, glycolaldehyde dehydrogenase, glycolate oxidase, and malate synthase. This series of enzymes will turn the ethylene glycol, released from the breakdown of PET, into malate which can be used by the cell as a carbon source via the citric acid cycle. <br><br> | + | <p>For our project, we have designed a plasmid that secretes MHETase and the double mutant PETase to increase the rate at which PET is degraded compared to the previous PETase sequence. We inserted the plasmid into an <em>E. coli </em>MG1655 strain. Because of the toxicity of ethylene glycol, a second plasmid was designed to allow the bacteria to break down the ethylene glycol and utilize its products as a carbon source. These enzymes include glycolaldehyde reductase, glycolaldehyde dehydrogenase, glycolate oxidase, and malate synthase. This series of enzymes will turn the ethylene glycol, released from the breakdown of PET, into malate which can be used by the cell as a carbon source via the citric acid cycle. <br><br> |
− | Our E. coli will degrade PET into terephthalic acid (TPA) and ethylene glycol (EG) utilizing the PETase and MHETase enzymes, as shown in Figure 1 below. </p> | + | Our <em>E. coli</em> will degrade PET into terephthalic acid (TPA) and ethylene glycol (EG) utilizing the PETase and MHETase enzymes, as shown in Figure 1 below. </p> |
<center> | <center> | ||
<img src = "https://static.igem.org/mediawiki/2018/9/9d/T--RHIT--design1.jpg" style="width:80%"> | <img src = "https://static.igem.org/mediawiki/2018/9/9d/T--RHIT--design1.jpg" style="width:80%"> | ||
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− | <p>Terephthalic acid is recyclable when purified and is relatively nontoxic. Ethylene glycol, on the other hand, is extremely toxic to humans, although it has been proven to be safe for E. coli cells. | + | <p>Terephthalic acid is recyclable when purified and is relatively nontoxic. Ethylene glycol, on the other hand, is extremely toxic to humans, although it has been proven to be safe for <em>E. coli </em>cells. |
<br><br> | <br><br> | ||
− | The E. coli will then take the ethylene glycol and utilize it as a carbon source using glycolaldehyde reductase, glycolaldehyde dehydrogenase, glycolate oxidase, and malate synthase. This will result in intermediates of glycolaldehyde, glycolate, and glyoxylate, as well as the product of malate which will be used in the citric acid cycle, as shown in Figure 2 below. </p> | + | The <em>E. coli</em> will then take the ethylene glycol and utilize it as a carbon source using glycolaldehyde reductase, glycolaldehyde dehydrogenase, glycolate oxidase, and malate synthase. This will result in intermediates of glycolaldehyde, glycolate, and glyoxylate, as well as the product of malate which will be used in the citric acid cycle, as shown in Figure 2 below. </p> |
<center> | <center> | ||
<img src = "https://static.igem.org/mediawiki/2018/3/30/T--RHIT--design2_.jpg"> | <img src = "https://static.igem.org/mediawiki/2018/3/30/T--RHIT--design2_.jpg"> |
Revision as of 19:22, 3 August 2018