Latest revision as of 02:48, 4 October 2012

Team:UC Davis - 2012.igem.org

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Project Overview

Environment-friendly plastic degradation has been a challenging and daunting task for our society. Plastic, one of the greatest discoveries of the 20th century is notoriously difficult to degrade or recycle in an efficient way. The overutilization of plastic and lack of integrated methods for its recycling has created major environmental threats that we are currently facing and will continue at a greater magnitude in the future. The Pacific Gyre patch, 3.5 million tons of floating trash that consists of nets, bottles, bags, among other more obscure items is such an example [2]. Plastic poses a serious threat to land and marine life, as animals often mistake it for food, or they consume byproducts that result from its partial degradation and are often toxic to them. Interestingly, collisions with floating or submerged waste objects has caused 269 boating accidents, resulting in 15 deaths, 116 injuries and 3 million dollars in damage, although this numbers are trivial when compared to the billion dollar catastrophes and uncertain future that this environmental hazard imposes to our society [3]. Furthermore, "It costs the state of California an estimated $72 million per year to collect and dispose of one-time use disposable cups and bags. In addition, it costs California an estimated $52.2 million per year to attempt to keep our beaches clean. In total, the current annual costs to public agencies for litter prevention, cleanup, and disposal is $375.2 million" [2].

Polyethylene terephthalate (PET) is a commonly used plastic due to its durability and molecular stability, but its high molecular weight and hydrophobicity also make it one of the hardest plastics to degrade. Although the PET recycling rate has increased from 7% to 30% in the last few years, a majority of the plastic is still dumped in landfills and continues to pollute the environment [1]. Most plastics are made from petroleum, oil or natural gas, and a variety of chemicals that are toxic to humans (including ethylene glycol). Phthalates and Bisphenol-A (BPA) are the most common types of chemicals used to make plastic materials. Phthalates have been known to cause diseases related to cancer or hormonal imbalances. BPA is often known to leak from bottles and migrate into liquids and foods it comes into contact with. Within the landfills, rainwater can trickle through the trash, creating toxic leachate. The leachate carries microorganisms and toxic chemicals into drinking water sources [2]. Instead of approaching the problem by reducing the amount of input coming in, most firms are simply increasing the amount of bioplastic. These “biobottles” are only 30% biodegradable, meaning that there is still an influx of non-biodegradable plastic coming into our environment [4]. Currently, the processes to recycle these plastics are churning out energy, water, and greenhouse gases, creating a process that is more wasteful than sustainable.

This year, the UC Davis IGEM team aspires to use synthetic biology techniques to create a microbial strain that has the capacity to metabolize PET and degrade it to non-toxic compounds. To achieve this outcome, we utilize targeted bioengineering and directed evolution techniques on the E. coli MG1655 strain, currently one of the most well-studied organisms. More specifically, the goals of our project are:

1. Targeted bioengineering of E. coli MG1655 to utilize PET as a carbon source. We seek to this by introducing a couple different modules to the strain. The first module encodes a cutinase gene which has been found to degrade PET into ethylene glycol and terephthalic acid. The second encodes the first two enzymes in a pathway that has been found to feed ethylene glycol into the TCA cycle.

2. Rational protein engineering to increase the enzymatic activity of cutinase to degrade PET.

3. Directed evolution of the ethylene glycol degradation pathway to increase its efficiency of ethylene glycol metabolism in E. coli cultures. Reintroduction of the glycolaldehyde reductase and glycolaldehyde dehydrogenase enzymes in various constructs to MG1655 and E-15 EG3 to increase the ethylene glycol consumption beyond E-15 EG3’s original capabilities.

References

1. Mazzoni, Mary. "PET Bottle Recycling Tops 1.5 Billion Pounds." Earth911. N.p., 13 Oct. 2011. Web. 4 Aug. 2012.
http://earth911.com/news/2011/10/13/pet-bottle-recycling-tops-1-5-billion-pounds/
2. "Pollution Prevention." Save Our Shores. Earth Resources, n.d. Web. 28 July 2012.
http://www.saveourshores.org/what-we-do/pollution-prevention.php
3. "Environmental Impact of The Garbage Patch." Environmentalimpact. University of Maine, Farmington, n.d. Web. 12 Aug. 2012.
http://students.umf.maine.edu/kevin.p.leary/public.www/environmentalimpact.html
4. Ghosh, Labonita. "Green Drive Makes Coca-Cola & Pepsi See Red Once Again." The Economic Times. Bennett, Coleman & Co. Ltd., 23 Feb. 2012. Web. 05 Aug. 2012.
http://articles.economictimes.indiatimes.com/2012-02-23/news/31091279_1_coca-cola-bottle-plant-bottle-carbon-footprint

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-        <ul>
+     <li ><a target="new" href="https://2012.igem.org/" title="Back to iGEM">iGEM</a>
-        <li class="groupb"><a href="https://2012.igem.org/" title="Back to iGEM">
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-		<img src="http://img.photobucket.com/albums/v26/bluemelon/main-button_09.jpg"></a></li>
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+          <ul>
+            <li ><a href="https://2012.igem.org/Team:UC_Davis/Data/Cutinase_Activity" title="Data">Cutinase Activity</a></li>
+            <li ><a href="https://2012.igem.org/Team:UC_Davis/Data/Ethylene_Glycol"
+title="Data">Ethylene Glycol</a></li>
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+title="Data">Modeling</a></li>
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+          <ul>
+            <li ><a href="https://2012.igem.org/Team:UC_Davis/Notebook">Notebook</a></li>
+            <li ><a href="https://2012.igem.org/Team:UC_Davis/Notebook/Protocols ">Protocols</a></li>
+            <li ><a href="https://2012.igem.org/Team:UC_Davis/Notebook/Gallery">Gallery</a></li>
+          </ul>
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-         <li class="groupb"><a href="https://2012.igem.org/Team:UC_Davis/Safety" title="Safety">
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-		<img src="https://static.igem.org/mediawiki/2012/d/df/UCDMain-button_04selected.jpg"></a></li>
+          <ul>
+            <li ><a href="https://2012.igem.org/Team:UC_Davis/Project">Project Overview</a></li>
+            <li ><a href="https://2012.igem.org/Team:UC_Davis/Project/Catalyst">Module Engineering</a></li>
+            <li ><a href="https://2012.igem.org/Team:UC_Davis/Project/Protein_Engineering">Protein Engineering</a></li>
+            <li ><a title="https://2012.igem.org/Team:UC_Davis/Project/Strain">Chassis Engineering </a>
+	   	<ul>
+                <li><a href="https://2012.igem.org/Team:UC_Davis/Project/Strain">Background</a></li>
+	        <li><a href="https://2012.igem.org/Team:UC_Davis/Project/Directed_Evolution">Directed Evolution</a></li>
+                <li><a href="https://2012.igem.org/Team:UC_Davis/Project/Our_Strain">Rational Engineering </a></li>
+		</ul>
+</li>
+          </ul>
+        </li>
+        <li ><a href="https://2012.igem.org/Team:UC_Davis/Team" title="Team">Team</a></li>
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-<p><!--Tell us more about your project.  Give us background.  Use this is the abstract of your project.  Be descriptive but concise (1-2 paragraphs)-->
-Polyethylene terephthalate (PET) is a commonly used plastic due to its durability and molecular stability, but its high molecular weight and hydrophobicity also make it one of the hardest plastics to degrade. PET is used in beverage containers, oven food trays, and video tapes. Although the PET recycling rate has increased from 7% to 30% in the last few years, a majority of the plastic is still dumped in landfills and continues to pollute the environment. Within the landfills, rainwater can trickle through the trash, creating toxic leachate. The leachate carries microorganisms and toxic chemicals into drinking water sources, making its impact go beyond the landfill. Plastic pollution is not isolated to landfills; plastic debris can wash up on shores and tons of plastic trash build up in the ocean. Marine animals can mistake the plastic debris as food, but since they cannot digest plastic they can starve. Plastic six pack rings can also choke animals by closing off the airway.
+<div id="sponsorbox" class="sponsorfloat">
+<center>
+<h2>Our Sponsors</h2>
+<a href="http://www.novozymes.com/en/Pages/default.aspx" target="_blank"><img src="https://static.igem.org/mediawiki/2011/2/21/UCD_Novozymes-logo.jpg" width="200"></a>
+</center>
-We hope to achieve biodegradation of PET by manipulating E. coli to secrete a key enzyme called cutinase, a lipolytic and esterolytic enzyme. Degradation of PET creates two by-products, one of which is a mildly toxic compound called ethylene glycol. Ethylene glycol will be broken down by two enzymes (glycolaldehyde reductase and glycolaldehyde dehydrogenase) into glycolate, a glycolysis intermediate. Aside from ethylene glycol, another byproduct of PET degradation is terephthalic acid (TPA). It is a harmless substance that can be reused for PET production. We will not be focusing on turning TPA into a more useful material. However, we do know that a certain strain of E. coli is able to convert TPA into polyhydroxyalkanoates (PHA), a kind of biodegradable plastic.
+<center>
+<a href="http://engineering.ucdavis.edu/" target="_blank"><img src="https://static.igem.org/mediawiki/2011/f/f6/UCD_CoE.png" width="200" height="40"></a>
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+<a href="http://biosci.ucdavis.edu/index_js.html" target="_blank"><img src="https://static.igem.org/mediawiki/2011/b/b1/UCD_biosci_sponsor.jpg" width="200" height="90"></a>
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+<a href="http://www.genomecenter.ucdavis.edu/" target="_blank"><img src="https://static.igem.org/mediawiki/2011/1/1b/UCD_Genome_center_sponsor.jpg" width="200" height="60"></a>
+</center>
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+" width="200"></a>
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+" width="200"></a>
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+" width="200"></a>
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-Even if the E. coli could successfully degrade a majority of the plastic, the strain would face intense competition if we were to try and implement is use in a landfill. E. coli will probably require a controlled, suitable environment to thrive in. One way to ensure this control would be to use a bioreactor.
-</p>
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-You are provided with this team page template with which to start the iGEM season.  You may choose to personalize it to fit your team but keep the same "look." Or you may choose to take your team wiki to a different level and design your own wiki.  You can find some examples.
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+<!-- div id="myleftbox" style="width:625px; float:left;" -->
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+<div id="myleftbox"  class="smallbox">
+<h1>Project Overview</h1>
+<article>
+Environment-friendly plastic degradation has been a challenging and daunting task for our society. Plastic, one of the greatest discoveries of the 20th century is notoriously difficult to degrade or recycle in an efficient way. The overutilization of plastic and lack of integrated methods for its recycling has created major environmental threats that we are currently facing and will continue at a greater magnitude in the future. The Pacific Gyre patch, 3.5 million tons of floating trash that consists of nets, bottles, bags, among other more obscure items is such an example [2]. Plastic poses a serious threat to land and marine life, as animals often mistake it for food, or they consume byproducts that result from its partial degradation and are often toxic to them. Interestingly, collisions with floating or submerged waste objects has caused 269 boating accidents, resulting in 15 deaths, 116 injuries and 3 million dollars in damage, although this numbers are trivial when compared to the billion dollar catastrophes and uncertain future that this environmental hazard imposes to our society [3]. Furthermore, "It costs the state of California an estimated $72 million per year to collect and dispose of one-time use disposable cups and bags. In addition, it costs California an estimated $52.2 million per year to attempt to keep our beaches clean. In total, the current annual costs to public agencies for litter prevention, cleanup, and disposal is $375.2 million" [2].
+<br>
+<center><img src="https://static.igem.org/mediawiki/2012/a/ad/Landfilllandscape.jpg"></center><br>Polyethylene terephthalate (PET) is a commonly used plastic due to its durability and molecular stability, but its high molecular weight and hydrophobicity also make it one of the hardest plastics to degrade. Although the PET recycling rate has increased from 7% to 30% in the last few years, a majority of the plastic is still dumped in landfills and continues to pollute the environment [1]. Most plastics are made from petroleum, oil or natural gas, and a variety of chemicals that are toxic to humans (including ethylene glycol). Phthalates and Bisphenol-A (BPA) are the most common types of chemicals used to make plastic materials. Phthalates have been known to cause diseases related to cancer or hormonal imbalances. BPA is often known to leak from bottles and migrate into liquids and foods it comes into contact with. Within the landfills, rainwater can trickle through the trash, creating toxic leachate. The leachate carries microorganisms and toxic chemicals into drinking water sources [2]. Instead of approaching the problem by reducing the amount of input coming in, most firms are simply increasing the amount of bioplastic. These “biobottles” are only 30% biodegradable, meaning that there is still an influx of non-biodegradable plastic coming into our environment [4]. Currently, the processes to recycle these plastics are churning out energy, water, and greenhouse gases, creating a process that is more wasteful than sustainable.
+<br><br><center><img src="https://static.igem.org/mediawiki/2012/a/a1/Saharsaharsahar.JPG" ></center><br>This year, the UC Davis IGEM team aspires to use synthetic biology techniques to create a microbial strain that has the capacity to metabolize PET and degrade it to non-toxic compounds.  To achieve this outcome, we utilize targeted bioengineering and directed evolution techniques on the E. coli MG1655 strain, currently one of the most well-studied organisms. More specifically, the goals of our project are:
+<br><br>1. Targeted bioengineering of <i>E. coli</i> MG1655 to utilize PET as a carbon source. We seek to this by introducing a couple different modules to the strain. The first module encodes a cutinase gene which has been found to degrade PET into ethylene glycol and terephthalic acid. The second encodes the first two enzymes in a pathway that has been found to feed ethylene glycol into the TCA cycle.
+<br><br>2. Rational protein engineering to increase the enzymatic activity of cutinase to degrade PET.
+<br><br>3. Directed evolution of the ethylene glycol degradation pathway to increase its efficiency of ethylene glycol metabolism in E. coli cultures. Reintroduction of the glycolaldehyde reductase and glycolaldehyde dehydrogenase enzymes in various constructs to MG1655 and E-15 EG3 to increase the ethylene glycol consumption beyond E-15 EG3’s original capabilities.
+</article></div>
+<br>
+<div id="myleftbox"  class="smallbox"><h1> References </h1>
+<article>
+.	Mazzoni, Mary. "PET Bottle Recycling Tops 1.5 Billion Pounds." Earth911. N.p., 13 Oct. 2011. Web. 4 Aug. 2012. <br>http://earth911.com/news/2011/10/13/pet-bottle-recycling-tops-1-5-billion-pounds/
+<br>
+.	"Pollution Prevention." Save Our Shores. Earth Resources, n.d. Web. 28 July 2012. <br>http://www.saveourshores.org/what-we-do/pollution-prevention.php
+<br>
+.	"Environmental Impact of The Garbage Patch." Environmentalimpact. University of Maine, Farmington, n.d. Web. 12 Aug. 2012. <br>http://students.umf.maine.edu/kevin.p.leary/public.www/environmentalimpact.html
+<br>
+.      Ghosh, Labonita. "Green Drive Makes Coca-Cola & Pepsi See Red Once Again." The Economic Times. Bennett, Coleman & Co. Ltd., 23 Feb. 2012. Web. 05 Aug. 2012. <br>http://articles.economictimes.indiatimes.com/2012-02-23/news/31091279_1_coca-cola-bottle-plant-bottle-carbon-footprint
+</article>
 </div>
-<p><br />
-</p><p><br />
-</p>
-<table align="justify">
-<tr>
-<td>You can write a background of your team here.  Give us a background of your team, the members, etc.  Or tell us more about something of your choosing.
-</td><td><div class="thumb tright"><div class="thumbinner" style="width:182px;"><a href="https://2012.igem.org/wiki/index.php?title=Special:Upload&amp;wpDestFile=UC_Davis_logo.png" class="new" title="File:UC Davis logo.png">File:UC Davis logo.png</a>  <div class="thumbcaption">200px</div></div></div>
-</td></tr>
-<tr>
-<td>
-<p><i>Tell us more about your project.  Give us background.  Use this as the abstract of your project.  Be descriptive but concise (1-2 paragraphs)</i>
-</p>
-</td><td><div class="thumb tright"><div class="thumbinner" style="width:182px;"><a href="/wiki/index.php?title=Special:Upload&amp;wpDestFile=UC_Davis_team.png" class="new" title="File:UC Davis team.png">File:UC Davis team.png</a>  <div class="thumbcaption">Your team picture</div></div></div>
-</td></tr>
-<tr>
-<td>
-</td><td align="center"><strong class="selflink"> Team UC_Davis</strong>
-</td></tr></table>
-<p><br />
-</p>
-<table style="color:#1b2c8a;background-color:#0c6;" cellpadding="3" cellspacing="1" border="1" width="62%" align="center">
-<tr>
-<th align="center"><strong class="selflink">Home</strong>
-</th><th align="center"><a href="https://2012.igem.org/Team:UC_Davis/Team" title="Team:UC Davis/Team">Team</a>
-</th><th align="center"><a href="https://2012.igem.org/Team.cgi?year=2012&amp;team_name=UC_Davis" class="external text" rel="nofollow">Official Team Profile</a>
-</th><th align="center"><a href="https://2012.igem.org/Team:UC_Davis/Project" title="Team:UC Davis/Project">Project</a>
-</th><th align="center"><a href="https://2012.igem.org/Team:UC_Davis/Parts" title="Team:UC Davis/Parts">Parts Submitted to the Registry</a>
-</th><th align="center"><a href="https://2012.igem.org/Team:UC_Davis/Modeling" title="Team:UC Davis/Modeling">Modeling</a>
-</th><th align="center"><a href="https://2012.igem.org/Team:UC_Davis/Notebook" title="Team:UC Davis/Notebook">Notebook</a>
-</th><th align="center"><a href="https://2012.igem.org/Team:UC_Davis/Safety" title="Team:UC Davis/Safety">Safety</a>
-</th><th align="center"><a href="https://2012.igem.org/Team:UC_Davis/Attributions" title="Team:UC Davis/Attributions">Attributions</a>
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+style="text-indent:-15px;
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+href="https://2012.igem.org/Team:UC_Davis/Project/Protein_Engineering
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+style="text-indent:-15px;list-style-image:none;list-style-type:none;color:#000000
+ "><li><a style="color:#000000 "
+href="https://2012.igem.org/Team:UC_Davis/Data/Cutinase_Activity ">
+Cutinase Activity</a> </li><li><a style="color:#000000 "
+href="https://2012.igem.org/Team:UC_Davis/Data/Ethylene_Glycol ">
+Ethylene Glycol</a> </li><li><a style="color:#000000 "
+href="https://2012.igem.org/Team:UC_Davis/Data/Modeling ">
+Modeling</a> </li><li><a style="color:#000000 "
+href="https://2012.igem.org/Team:UC_Davis/Parts ">Parts</a></li> </ul>
+<li style="float:left ;margin:0 10px"><a
+href="https://2012.igem.org/Team:UC_Davis/Attributions ">
+<p>Attribution </p></a><a style="color:#000000 "
+href="https://2012.igem.org/Team:UC_Davis/Attributions ">
+Attribution</a></li>
+<li style="float:left ;margin:0 10px"><a
+href="https://2012.igem.org/Main_Page "> <p>iGEM </p></a><ul
+style="text-indent:-15px;list-style-image:none;list-style-type:none;color:#000000
+ "><li><a style="color:#000000 " href="https://2012.igem.org/Main_Page
+">Main iGEM</a> </li><li><a style="color:#000000 "
+href="https://2012.igem.org/Team:UC_Davis/Criteria "> Criteria</a>
+</li><li><a style="color:#000000 "
+href="https://2012.igem.org/Team:UC_Davis/Human_Practices ">Human
+Practices</a></li> </ul>
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