Difference between revisions of "Team:RHIT/Description"

 
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  <img id="dsc" src = "https://static.igem.org/mediawiki/2018/4/47/T--RHIT--petri.jpg" style="width:140px">
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      <h6 style="font-size:100%" id="dsct"> Description </h6>
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<a href="https://2018.igem.org/Team:RHIT/Design">
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  <img id="des" src="https://static.igem.org/mediawiki/2018/d/da/T--RHIT--petrihover.jpg" style="width:140px">
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      <h6 style="font-size:100%" id="dest"> Design </h6> </a>
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<a href = "https://2018.igem.org/Team:RHIT/Experiments">
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  <img id="exp" src="https://static.igem.org/mediawiki/2018/d/da/T--RHIT--petrihover.jpg" style="width:140px">
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    <img class="p" id="exp" src="https://static.igem.org/mediawiki/2018/4/47/T--RHIT--petri.jpg" style="width:140px">
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<a href="https://2018.igem.org/Team:RHIT/Design">
 
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      <h6 style="font-size:100%" id="notet"> Notebook </h6></a>
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<a href="https://2018.igem.org/Team:RHIT/Results">  
 
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      <h6 style="font-size:100%" id="rest"> Results </h6></a>
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<a href="https://2018.igem.org/Team:RHIT/Achievements">  
 
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        <h6 style="font-size:100%" id="acht"> Achievements </h6></a>
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      <h6 style="font-size:100%" id="inlt"> InterLab </h6> </a>
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<a href="https://2018.igem.org/Team:RHIT/Attributions">
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<h6 style="font-size:100%" id="attrt"> Attributions </h6> </a>
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 +
 
 +
<div class = "column full_size">
 
<h5> The Plastic Problem </h5>
 
<h5> The Plastic Problem </h5>
−
<p> The Great Pacific Garbage Patch contains over 1.8 trillion pieces of plastic, and weighs over 100,000 tons. It covers an area over 1.6 million square kilometers- three times the area of France. The plastics that have made their way into the ocean will eventually break down into microplastics, and make their way into the lives of unsuspecting and unfortunate marine life. Studies on fish and other marine animals have shown that over 10% have plastics in their systems. </br> </br>
+
<p> We live in a plastic world. From our grocery bags and disposable bottles to straws and coffee cups, plastic has become an integrated part of our everyday lives. Since plastic production started in the 1950s, more than 6.9 billion tons of plastic have become waste. According to a study in 2017, only 9% of this waste has been recycled [1]. If these products never reached the recycling plant, where did 6.3 billion tons of plastic end up? Unfortunately, the plastics that are never recycled end up in landfills, on the sides of roads, and in our lakes, rivers, and oceans. </p>
 +
<center>
 +
<img src = "https://static.igem.org/mediawiki/2018/d/d8/T--RHIT--DescBottles.jpg" style="width:60%">
 +
</center>
 +
<center>
 +
PHOTOGRAPH BY RANDY OLSON
 +
</center>
 +
<br><br>
 +
<p>Because of its unique structure, plastic does not degrade. Instead, the plastic just breaks up into smaller and smaller pieces, eventually becoming microplastics. Smaller pieces of plastic and microplastics are more dangerous because they are easier for sealife to ingest. Because of the currents in the oceans, the plastic dumped in the ocean can travel around the world, leaving no place unaffected. As much as 15% of sand on certain beaches in Hawaii is made of microplastics [1].</p>
 +
<br><br>
  
−
Polyethylene terephthalate (PET) is a kind of plastic used in an innumerable amount of products from water bottles to personal care product packaging. Most of the plastic that makes its way to pollute the ocean and other ecosystems is made of PET. A study showed that over 6.3 billion metric tons of plastic waste had been created by humans, and only a sad 9% of that plastic has been recycled. A main contributing factor to this plastic problem is the large volume of PET plastic bottles produced and used in the world. 1 million plastic bottles are sold every minute, and most of them are thrown away instead of recycled.</p> </div>  
+
<h5> Previous Work </h5>
 +
<p>In March of 2016, an article was released about the discovery of a bacterium that could degrade PET, polyethylene terephthalate. Ideonella sakaiensis 201-F6 was found to contain a PET hydrolase and a MHET hydrolase, named PETase and MHETase respectively. PETase introduced on a PET film degraded PET into MHET, a monomer of the PET chain, along with minimal amounts of terephthalic acid. In combination with MHETase, the PET film was degraded into the final products of terephthalic acid and ethylene glycol. After 6 weeks, the PET film was almost completely degraded [2]. <br><br>  
  
 +
Two years later, in April of 2018, an article was released with extensive research on wild-type PETase and select PETase mutations. The W159H/S238F double mutant of PETase showed significant improvement in crystallinity reduction and product release over the wild-type in just 96 hours. The percent crystallinity change is a result of the pitting on the film caused by the PET degradation [3]. </p> <br>
 +
<center>
 +
<img src = "https://static.igem.org/mediawiki/2018/2/21/T--RHIT--DescGraph.jpg" style="width:50%">
 +
</center>
 +
<center> Figure 1. A comparison of the previous PETase sequence and the double-mutated sequence. Image included from the April 2018 article [3].
 +
</center>
 +
<br><br>
  
−
<div class = "column two_thirds_size">
+
<h5> Our Project and Inspiration </h5>
−
<h5> Our Goal </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 <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>  
−
<p>Researchers in Japan have recently found a bacteria, Ideonella sakaiensis, that eats PET plastic. PET can be broken down into ethylene glycol and terephthalic acid. We are taking the sequence for the enzyme that breaks down PET and putting it into an E. coli plasmid. Because of the toxicity of ethylene glycol, another sequence was created to let the bacteria obtain energy by breaking down the ethylene glycol and using it as the bacteria’s only carbon source. In the end, the bacteria will have transformed PET plastic into only terephthalic acid that can be recycled to be turned back into new plastics.
+
−
</br></br>
+
−
This year, Rose-Hulman’s Six Sigma club hosted a talk from ​Dr. Shikha Bhattacharyya, who discussed straw usage in the US and how to reduce it. After this, the school started offering biodegradable straws that were compostable. Although this is a great solution for plastic straws, there was still no solution to the plastic bottle problem. After researching the Ideonella sakaiensis, we realized that using bacteria was a promising way to tackle the plastic bottle problem. </p></div>
+
  
−
<div class = "column third_size">
+
Much of the inspiration for our project came from the many changes that have taken place on Rose-Hulman’s campus. Within the last few years, recycling and decreasing plastic waste have become important aspects of campus. Many of these changes are due to the Six Sigma class. Six Sigma does projects where the students collect data before and after an improvement phase. Some of their past projects have included collecting data on the amount of recyclables in the trash and the use of plastic straws on campus. Recycling areas were set up throughout the academic buildings, and the campus community was educated about what and how to recycle. The plastic straw project initiated a decline in plastic straw use around campus by offering a biodegradable alternative to plastic straws at the eateries on campus and selling reusable straws to students. Figure 2 and Figure 3 show data collected by the Six Sigma class during their recycling project. Figure 2 and 3 show data before and after the improvement phase. They show a noticeable change in the campus community [4]. </p> <br>
−
</br></br>
+
−
<img src = "https://static.igem.org/mediawiki/2018/3/30/T--RHIT--CorrectedPebble.jpg">
+
−
</div>
+
  
 +
<center>
 +
<img src = "https://static.igem.org/mediawiki/2018/c/cb/T--RHIT--descBefore.jpg" style="width:60%">
 +
</center>
 +
<center>Figure 2. Graphical summary of the percentage of recyclables data over the baseline period [4]. </center>
 +
<br><br>
 +
<center>
 +
<img src = "https://static.igem.org/mediawiki/2018/c/cd/T--RHIT--descAfter.jpg" style="width:60%">
 +
</center>
 +
<center>Figure 3. Graphical summary of the percentage of recyclables collected over the improvement period [4]. </center>
 +
<br><br>
  
−
</div>
 
−
<div class = "column full_size">
 
 
<h5> References </h5>
 
<h5> References </h5>
 
<ul>
 
<ul>
−
<li><a href = "https://www.theoceancleanup.com/great-pacific-garbage-patch/"> https://www.theoceancleanup.com/great-pacific-garbage-patch/</a> </li>
+
<li>[1] Parker, L. (2018). We Depend On Plastic. Now, We’re Drowning in It.. [online] Nationalgeographic.com. </li>
  
−
<li><a href = "http://garbagepatch.net/greatpacificoceangarbagepatchfacts/"> http://garbagepatch.net/greatpacificoceangarbagepatchfacts/ </a></li>
+
<li>[2] Yoshida, S., Hiraga, K., Takehana, T., Taniguchi, I., Yamaji, H., Maeda, Y., Toyohara, K., Miyamoto, K., Kimura, Y. and Oda, K. (2016). A bacterium that degrades and assimilates poly(ethylene terephthalate). Science, 351(6278), pp.1196-1199.</li>
  
−
<li><a href = "https://helpsavenature.com/great-pacific-garbage-patch-facts"> https://helpsavenature.com/great-pacific-garbage-patch-facts </a></li>
+
<li>[3] Austin, H., Allen, M., Donohoe, B., Rorrer, N., Kearns, F., Silveira, R., Pollard, B., Dominick, G., Duman, R., El Omari, K., Mykhaylyk, V., Wagner, A., Michener, W., Amore, A., Skaf, M., Crowley, M., Thorne, A., Johnson, C., Woodcock, H., McGeehan, J. and Beckham, G. (2018). Characterization and engineering of a plastic-degrading aromatic polyesterase. Proceedings of the National Academy of Sciences, 115(19), pp.E4350-E4357.</li>
  
−
<li><a href = "https://www.globalcitizen.org/en/content/9-facts-about-ocean-plastic/"> https://www.globalcitizen.org/en/content/9-facts-about-ocean-plastic/</a></li>
+
<li>[4] D. Evans and P. Olejnik, “Tightening Rose-Hulman's Wasteline: Using a Standard Operating
 +
Procedure to Reduce Trashed Recyclables on a College Campus,” rep.</li>
  
−
<li><a href = "https://news.nationalgeographic.com/2018/03/great-pacific-garbage-patch-plastics-environment/"> https://news.nationalgeographic.com/2018/03/great-pacific-garbage-patch-plastics-environment/</a></li>
 
 
</ul>
 
</ul>
 
</div>
 
</div>
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<div class = "clear extra_space"></div>
 
  
  
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<div class="column two_thirds_size">
 
−
<h3>What should this page contain?</h3>
 
−
<ul>
 
−
<li> A clear and concise description of your project.</li>
 
−
<li>A detailed explanation of why your team chose to work on this particular project.</li>
 
−
<li>References and sources to document your research.</li>
 
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<li>Use illustrations and other visual resources to explain your project.</li>
 
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</ul>
 
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</div>
 
  
−
<div class="column third_size" >
 
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<h3>Inspiration</h3>
 
−
<p>See how other teams have described and presented their projects: </p>
 
  
−
<ul>
 
−
<li><a href="https://2016.igem.org/Team:Imperial_College/Description">2016 Imperial College</a></li>
 
−
<li><a href="https://2016.igem.org/Team:Wageningen_UR/Description">2016 Wageningen UR</a></li>
 
−
<li><a href="https://2014.igem.org/Team:UC_Davis/Project_Overview"> 2014 UC Davis</a></li>
 
−
<li><a href="https://2014.igem.org/Team:SYSU-Software/Overview">2014 SYSU Software</a></li>
 
−
</ul>
 
−
</div>
 
  
  
−
<div class="column two_thirds_size" >
 
−
<h3>Advice on writing your Project Description</h3>
 
  
−
<p>
+
<div class = "column full_size">
−
We encourage you to put up a lot of information and content on your wiki, but we also encourage you to include summaries as much as possible. If you think of the sections in your project description as the sections in a publication, you should try to be concise, accurate, and unambiguous in your achievements.
+
−
</p>
+
  
 
</div>
 
</div>
−
 
−
<div class="column third_size">
 
−
<h3>References</h3>
 
−
<p>iGEM teams are encouraged to record references you use during the course of your research. They should be posted somewhere on your wiki so that judges and other visitors can see how you thought about your project and what works inspired you.</p>
 
−
 
−
</div>
 
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Latest revision as of 02:38, 13 October 2018




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patekm@rose-hulman.edu
5500 Wabash Ave.
Terre Haute, IN 47803

Description
Design
Experiments
Notebook
Results
Achievements
InterLab
Attributions
The Plastic Problem

We live in a plastic world. From our grocery bags and disposable bottles to straws and coffee cups, plastic has become an integrated part of our everyday lives. Since plastic production started in the 1950s, more than 6.9 billion tons of plastic have become waste. According to a study in 2017, only 9% of this waste has been recycled [1]. If these products never reached the recycling plant, where did 6.3 billion tons of plastic end up? Unfortunately, the plastics that are never recycled end up in landfills, on the sides of roads, and in our lakes, rivers, and oceans.

PHOTOGRAPH BY RANDY OLSON


Because of its unique structure, plastic does not degrade. Instead, the plastic just breaks up into smaller and smaller pieces, eventually becoming microplastics. Smaller pieces of plastic and microplastics are more dangerous because they are easier for sealife to ingest. Because of the currents in the oceans, the plastic dumped in the ocean can travel around the world, leaving no place unaffected. As much as 15% of sand on certain beaches in Hawaii is made of microplastics [1].



Previous Work

In March of 2016, an article was released about the discovery of a bacterium that could degrade PET, polyethylene terephthalate. Ideonella sakaiensis 201-F6 was found to contain a PET hydrolase and a MHET hydrolase, named PETase and MHETase respectively. PETase introduced on a PET film degraded PET into MHET, a monomer of the PET chain, along with minimal amounts of terephthalic acid. In combination with MHETase, the PET film was degraded into the final products of terephthalic acid and ethylene glycol. After 6 weeks, the PET film was almost completely degraded [2].

Two years later, in April of 2018, an article was released with extensive research on wild-type PETase and select PETase mutations. The W159H/S238F double mutant of PETase showed significant improvement in crystallinity reduction and product release over the wild-type in just 96 hours. The percent crystallinity change is a result of the pitting on the film caused by the PET degradation [3].


Figure 1. A comparison of the previous PETase sequence and the double-mutated sequence. Image included from the April 2018 article [3].


Our Project and Inspiration

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.

Much of the inspiration for our project came from the many changes that have taken place on Rose-Hulman’s campus. Within the last few years, recycling and decreasing plastic waste have become important aspects of campus. Many of these changes are due to the Six Sigma class. Six Sigma does projects where the students collect data before and after an improvement phase. Some of their past projects have included collecting data on the amount of recyclables in the trash and the use of plastic straws on campus. Recycling areas were set up throughout the academic buildings, and the campus community was educated about what and how to recycle. The plastic straw project initiated a decline in plastic straw use around campus by offering a biodegradable alternative to plastic straws at the eateries on campus and selling reusable straws to students. Figure 2 and Figure 3 show data collected by the Six Sigma class during their recycling project. Figure 2 and 3 show data before and after the improvement phase. They show a noticeable change in the campus community [4].


Figure 2. Graphical summary of the percentage of recyclables data over the baseline period [4].


Figure 3. Graphical summary of the percentage of recyclables collected over the improvement period [4].


References
  • [1] Parker, L. (2018). We Depend On Plastic. Now, We’re Drowning in It.. [online] Nationalgeographic.com.
  • [2] Yoshida, S., Hiraga, K., Takehana, T., Taniguchi, I., Yamaji, H., Maeda, Y., Toyohara, K., Miyamoto, K., Kimura, Y. and Oda, K. (2016). A bacterium that degrades and assimilates poly(ethylene terephthalate). Science, 351(6278), pp.1196-1199.
  • [3] Austin, H., Allen, M., Donohoe, B., Rorrer, N., Kearns, F., Silveira, R., Pollard, B., Dominick, G., Duman, R., El Omari, K., Mykhaylyk, V., Wagner, A., Michener, W., Amore, A., Skaf, M., Crowley, M., Thorne, A., Johnson, C., Woodcock, H., McGeehan, J. and Beckham, G. (2018). Characterization and engineering of a plastic-degrading aromatic polyesterase. Proceedings of the National Academy of Sciences, 115(19), pp.E4350-E4357.
  • [4] D. Evans and P. Olejnik, “Tightening Rose-Hulman's Wasteline: Using a Standard Operating Procedure to Reduce Trashed Recyclables on a College Campus,” rep.