Difference between revisions of "Team:OLS Canmore Canada/Demonstrate"

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   <h1 class="headertext">DESIGN</h1>
 
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<h1 class="title">The Summary</h1>
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<h1 class="title">The Design</h1>
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The project will use synthetic biology to create a novel fusion protein that can specifically bio-tag polyethylene terephthalate (PET) plastic, so that it can be sorted and recycled correctly. Synthetic biology is efficient, cost effective, and specific. The proteins, which are produced via a bacterial chassis called Bacillus subtilis, are created efficiently and at low cost. These proteins also provide high specificity due to a specific 3-dimensional shape that adheres selectively to PET polymers.  The 4 constructs that we have designed, with the help of our mentors and previous iGem teams, include:
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In recent years, the issue of plastic pollution has become an overwhelming global crisis. Only 5% of all plastics are recycled and the rest ends up in landfills or oceans. When looking for a solution to this problem, the Design Thinking methodology learned at the Berkeley Program was applied. In the engagement with recycling stakeholders, the OLS SynBio team discovered that the issue is not the recycling of plastic, but instead the inefficient sorting of these plastics.</p>
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<p>To further understand this issue, the team participated in many community outreach events. OLS Synbio consulted with Simon Robbins, the corporate manager of a local recycling plant, who provided guidance and insight of how the recycling cycle works. OLS SynBio also met with Peter Duck, the zero waste manager for the town of Canmore. Lastly, the team went to the Alberta Recycling Conference to learn more about how plastics are recycled in the community, and how big the team’s contribution would be. Stakeholder feedback helped to pivot and refine the project. </p>
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<li>a polyethylene terephthalate hydrolase (PET-ase) fused to a red fluorescent protein, (or RFP) called mCherry, which give the protein its <b>colour</b> aspect. </li>
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<li>a hydrophobin called BslA,</li>
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<li>a PET-ase without the RFP, and </li>
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<tr><td><img  width="100%"src="image1.png"></td></tr>
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<li>a BslA without RFP.</li>
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<tr><td class="imagecaptiontext">Some members of our team at the Canmore Sorting Facility.</td></tr>
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<h1 class="subtitle">The Subtitle</h1>
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A LipA secretion tag is added to each construct to signal the bacteria to secrete the proteins out of the cell for easier purification. We chose to use this Bacillus over E. coli because of its natural ability to produce hydrophobins, and because it is better at secreting proteins than other bacteria.  Bacillus is also naturally occurring in the environment, and has reduced risk for environmental contamination concerns.
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</p>
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<br>
  
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<p>The project will use synthetic biology to create a novel fusion protein that can specifically bio-tag polyethylene terephthalate (PET) plastic, so that it can be sorted and recycled correctly. The project involves two proteins, a polyethylene terephthalate hydrolase (PETase) and a hydrophobin called BsIA, that is produced by a bacterium chassis called Bacillus subtilis. The PETase protein naturally binds to PET and would be paired with a red fluorescent protein called mCherry to visually indicate when the protein has adhered. The hydrophobin is “water-fearing”, therefore it will bind to anything, but for this project, it will help to adhere the PETase specifically to PET plastic. The project plan is to experiment with the use of both proteins, together and independently. If successful, the bio-tag would be proof of concept for a novel technology that can be implement easily in existing recycling facilities.
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<p>
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The PET-ase is an enzyme that naturally binds to PET plastic, and the mCherry RFP it is paired with will visually indicate when the protein has adhered. The hydrophobin is “water-fearing” and will therefore bind to several surfaces. However, for this project, it will be used to help adhere the PET-ase specifically to PET plastic. We are using the four proteins in combination with each other and test their effectiveness at tagging PET plastic.  
 
</p>
 
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<tr><td><img  width="100%" src="https://static.igem.org/mediawiki/2018/d/d9/T--OLS_Canmore_Canada--prototypetext.svg"></td></tr>
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<tr><td class="imagecaptiontext">Implemented prototype, what could be seen in a sorting facility.</td></tr>
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<h1 class="subtitle">Machine Prototype</h1>
 
<p>
 
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Before incorporating it into the recycling facility, the protein would be isolated and purified, and the team will run numerous proof-of-concept assays. The next step in the project is prototyping. The team has explored a prototype which would use a streamlined linear process that involves both existing technology and new robotics to effectively sort plastics. Early business modelling suggests that this project is desirable by people, feasible with technology and viable as a business. 
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With our constructs design in place, we had to design a way of using them in a real life situation.  Drawing on our experiences visiting real sorting facilities, and using the feedback and insights gained from the people working in this industry,  we have designed a prototype using existing technology to adapt to our solution. A simplified description of our prototype includes the following steps:
 
</p>
 
</p>
 
<br>
 
<br>
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<p>In summary, the OLS SynBio team is creating a novel protein bio-tag that will adhere selectively to PET plastics.  This product has the potential to revolutionize the recycling industry, and reduce the current practice of landfilling poorly sorted plastics. This will create a truly circular life cycle for plastic products.</p>
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<ol style="margin-bottom: 20vh;" class="standard">
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<li>Incoming, unsorted plastics move along a conveyor belt and pass through a bath of our purified protein bio-tag. </li>
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<li>Our bio-tag selectively adheres only to PET plastics. </li>
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<li>Next all plastics will pass through a wash or rinse.  The bio-tag is removed from any non-PET plastics.</li>
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<li>An optical scanner detects the fluorescent signature of mCherry on the PET plastics, and will separate it from the rest of the plastic. </li>
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<li>In future, similar bio-tags can be developed to selectively mark all other recyclable plastics using similar design principles.</li>
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</ol>
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Revision as of 03:45, 17 October 2018

DESIGN

The Design

The project will use synthetic biology to create a novel fusion protein that can specifically bio-tag polyethylene terephthalate (PET) plastic, so that it can be sorted and recycled correctly. Synthetic biology is efficient, cost effective, and specific. The proteins, which are produced via a bacterial chassis called Bacillus subtilis, are created efficiently and at low cost. These proteins also provide high specificity due to a specific 3-dimensional shape that adheres selectively to PET polymers. The 4 constructs that we have designed, with the help of our mentors and previous iGem teams, include:


  • a polyethylene terephthalate hydrolase (PET-ase) fused to a red fluorescent protein, (or RFP) called mCherry, which give the protein its colour aspect.
  • a hydrophobin called BslA,
  • a PET-ase without the RFP, and
  • a BslA without RFP.

A LipA secretion tag is added to each construct to signal the bacteria to secrete the proteins out of the cell for easier purification. We chose to use this Bacillus over E. coli because of its natural ability to produce hydrophobins, and because it is better at secreting proteins than other bacteria. Bacillus is also naturally occurring in the environment, and has reduced risk for environmental contamination concerns.


The PET-ase is an enzyme that naturally binds to PET plastic, and the mCherry RFP it is paired with will visually indicate when the protein has adhered. The hydrophobin is “water-fearing” and will therefore bind to several surfaces. However, for this project, it will be used to help adhere the PET-ase specifically to PET plastic. We are using the four proteins in combination with each other and test their effectiveness at tagging PET plastic.

Implemented prototype, what could be seen in a sorting facility.

Machine Prototype

With our constructs design in place, we had to design a way of using them in a real life situation. Drawing on our experiences visiting real sorting facilities, and using the feedback and insights gained from the people working in this industry, we have designed a prototype using existing technology to adapt to our solution. A simplified description of our prototype includes the following steps:


  1. Incoming, unsorted plastics move along a conveyor belt and pass through a bath of our purified protein bio-tag.
  2. Our bio-tag selectively adheres only to PET plastics.
  3. Next all plastics will pass through a wash or rinse. The bio-tag is removed from any non-PET plastics.
  4. An optical scanner detects the fluorescent signature of mCherry on the PET plastics, and will separate it from the rest of the plastic.
  5. In future, similar bio-tags can be developed to selectively mark all other recyclable plastics using similar design principles.