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

 
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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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With our solution and constructs in place, the OLS SynBio team has focussed intently on implementing our project into the real world. We had many questions about how we were going to introduce our idea into sorting facilities. Over many months of discussion, research, and brainstorming, we designed a prototype model. While brainstorming ideas for our prototype and how we were going to implement it into today’s society, we discussed many conflicts, such as the safety of our solution entering the recycling facilities, how it was going to affect the workers, and if facilities even were interested in incorporating our system. In the future, we hope to place our system into a sorting facility in order to increase the efficiency of sorting and provide a solution to the global crisis of plastic in our environment. </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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Before brainstorming a biological marker to tag on to the plastic, we researched existing forms of plastic sorting. These techniques include manual sorting, which is dangerous, and expensive sorting technologies, such as laser scanners. With these types being the main sorting method, we created a bio-tag and a prototype that goes alongside it, which implements existing, inexpensive technology. We started off with many design aspects when creating our prototype. We have been adapting existing technology to work best with the bio-tag. We collaborated with the OLS High School Robotics team to help improve the feasibility of our prototype. The optical scanner that detects the mCherry on the PET plastics was put to the test by the robotics team. As a result of all their hard work and engineering skills, they have successfully manufactured a robot that can detect certain colours. One major issue surrounding the use of a biological marker is that it can raise some ethical concerns regarding the safety of humans and the environment. To address this, the bio-tag is going to be a purified protein. This means that no live cells are going to come in contact with the plastic. With the bio-tag created, many people have a similar main concern on how much of it was going to come in contact with the environment, workers, and facilities. Our team decided that a bath of the protein was the best option to eliminate an aerosol effect that a spray or waterfall could create. By putting the plastics in the bath, the chance of contamination of the biological solution within the recycling facilities is decreased. We also discussed the importance of a water rinse after the plastic goes through the bio-tag solution. This ensures that any proteins that are not bound to the PET plastic would not impact the results and readings of the optical scanner. After the water rinse, only the bio-tag should be left on the plastics. This prototype is a closed system; thus further reducing the chance of contamination in the environment.  
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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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<li>a BslA without RFP.</li>
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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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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.
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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>
 
<tr><td><img  width="100%" src="https://static.igem.org/mediawiki/2018/d/d9/T--OLS_Canmore_Canada--prototypetext.svg"></td></tr>
 
<tr><td class="imagecaptiontext">Implemented prototype, what could be seen in a sorting facility.</td></tr>
 
<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>
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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:
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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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<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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<li>Our bio-tag selectively adheres only to PET plastics. </li>
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<tr><td class="imagecaptiontext">Implemented prototype, what could be seen in a sorting facility.</td></tr>
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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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Latest revision as of 02:41, 18 October 2018

DEMONSTRATE

Demonstrate

With our solution and constructs in place, the OLS SynBio team has focussed intently on implementing our project into the real world. We had many questions about how we were going to introduce our idea into sorting facilities. Over many months of discussion, research, and brainstorming, we designed a prototype model. While brainstorming ideas for our prototype and how we were going to implement it into today’s society, we discussed many conflicts, such as the safety of our solution entering the recycling facilities, how it was going to affect the workers, and if facilities even were interested in incorporating our system. In the future, we hope to place our system into a sorting facility in order to increase the efficiency of sorting and provide a solution to the global crisis of plastic in our environment.


Before brainstorming a biological marker to tag on to the plastic, we researched existing forms of plastic sorting. These techniques include manual sorting, which is dangerous, and expensive sorting technologies, such as laser scanners. With these types being the main sorting method, we created a bio-tag and a prototype that goes alongside it, which implements existing, inexpensive technology. We started off with many design aspects when creating our prototype. We have been adapting existing technology to work best with the bio-tag. We collaborated with the OLS High School Robotics team to help improve the feasibility of our prototype. The optical scanner that detects the mCherry on the PET plastics was put to the test by the robotics team. As a result of all their hard work and engineering skills, they have successfully manufactured a robot that can detect certain colours. One major issue surrounding the use of a biological marker is that it can raise some ethical concerns regarding the safety of humans and the environment. To address this, the bio-tag is going to be a purified protein. This means that no live cells are going to come in contact with the plastic. With the bio-tag created, many people have a similar main concern on how much of it was going to come in contact with the environment, workers, and facilities. Our team decided that a bath of the protein was the best option to eliminate an aerosol effect that a spray or waterfall could create. By putting the plastics in the bath, the chance of contamination of the biological solution within the recycling facilities is decreased. We also discussed the importance of a water rinse after the plastic goes through the bio-tag solution. This ensures that any proteins that are not bound to the PET plastic would not impact the results and readings of the optical scanner. After the water rinse, only the bio-tag should be left on the plastics. This prototype is a closed system; thus further reducing the chance of contamination in the environment.


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

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