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| - | <h1>Data</h1> | + | <h1>Cutinase Activity</h1> |
| | <article> | | <article> |
| - | The accumulation of plastic products poses a hazard to the environment, as well as humans, through drinking water contamination. This threat led us to develop a degradation pathway to turn the polyethylene terephthalate into different substrates. We produce terephthalic acid and ethylene glycol. Ethylene glycol is a moderately toxic substance, which is oxidized to glycolic acid. The glycolic acid is further oxidized to oxalic acid – a toxic substance that affects the central nervous system via the liver. However, in the environment, the ethylene glycol will be degraded by hydroxyl radicals and in sewage sludge, it is readily biodegradable. Because ethylene glycol must be ingested to pose a problem, researchers take extra precaution to make sure there are no splashes of ethylene glycol in the laboratory and the wastes will be disposed of in the appropriate hazardous waste receptacles. Ethylene glycol can also be a mild irritant if it comes in contact with the skin or if it is inhaled, so researchers wear eye protection as well as gloves and lab coats, and always work with ethylene glycol in the confine of a fume hood. Also, in our constructs, we have produced enzymes that will degrade ethylene glycol into glycoaldehyde and then glycolate. The glycolate has the potential to be turned in to oxaloacetate, a metabolic intermediate. In the environment, ethylene glycol can potentially be toxic within waterways, however the team made sure to dispose of ethylene glycol in a responsible way.
| + | As of now, through our p-nitrophenyl butyrate assays we have gathered enough data to determine that our LC-Cutinase part (<a href="http://partsregistry.org/wiki/index.php?title=Part:BBa_K936000">BBa_K936000</a>) exhibits its intended function as an esterase. Unfortunately it has been much more difficult to characterize than expected as we have been unable to purify the protein successfully. |
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| - | <p>Toxicity of EG Graph for MG1655 and DH5a</p>
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| - | We observed that EG is non-toxic to both DH5a and MG1655 cells, as evident from the growth of the two strains. Both strains were exposed to LB media containing varying amounts of EG, ranging from 0 mM to 150 mM.
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| - | <br><br><center> | + | </article></div> |
| - | <a href="https://static.igem.org/mediawiki/2012/d/d3/UCD_DH5a_data_1_large.jpg" class="lightbox">
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| - | <img src="https://static.igem.org/mediawiki/2012/2/2b/UCD_DH5a_data_1.jpg" width="300" align="left"></a>
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| - | <a href="https://static.igem.org/mediawiki/2012/7/7e/UCD_MG1655_data_1_large.jpg" class="lightbox">
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| - | <img src="https://static.igem.org/mediawiki/2012/4/43/UCD_MG1655_data_1.jpg" width="300" align="right"></a>
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| - | </center>
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| - | <br><p>Spain in EG Graph</p>
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| - | Knowing that K-12 Strain E-15 EG3 utilizes ethylene glycol, we devised an experiment to test the optimal amount of ethylene glycol the strain would thrive in. From the graph, we deduced that 30 mM of ethylene glycol showed the highest growth rate, which also matched previous data provided in the referenced paper [1].
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| - | <a href="https://static.igem.org/mediawiki/2012/3/3b/UCD_EG_large_1.jpg" class="lightbox"><img src="https://static.igem.org/mediawiki/2012/6/67/UCD_EG_1.jpg"></a>
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| | <h1>References</h1> | | <h1>References</h1> |
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| - | 1. Boronat, Albert, Caballero, Estrella, and Juan Aguilar. “Experimental Evolution of a Metabolic Pathway for Ethylene Glycol Utilization by Escherichia coli.” Journal of Bacteriology, Vol. 153 No. 1, pp. 134-139, January 1983.)
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| | </article></div> | | </article></div> |
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