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

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<h1 class="title">The Design</h1>
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<h1 class="title">Precautions</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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The OLS igem Team has taken many precautions to ensure a safe lab environment, and the environmental safety of our new constructs. Some of the steps that have been taken to further ensure safety are:
 
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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>In the design of the construct our team chose to use a lab strain of Bacillus subtilis, as it is a naturally occurring bacteria found in dirt and plants, so it is not harmful to the environment. </li>
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<li>a hydrophobin called BslA,</li>
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<li></li>
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<li>a PET-ase without the RFP, and </li>
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<li>Our construct is intended for use as a protein bio-tag manufacturing system.  The bio-tag protein would be isolated and purified for use, ensuring no live bacteria enter a sorting or recycling facility at any time.</li>
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<li>a BslA without RFP.</li>
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<li>In the process of applying our bio tag in the real world, the plastics will be bathed, rather than sprayed with the protein, to avoid airborne proteins and potential contamination. </li>
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<li>Once plastic has been bio-tagged and sorted, it can now get recycled. In the recycling process plastic is shredded and melted down. The heat that is needed to melt the plastic will denature the proteins to stop potential environmental contamination. With future trials, the protein’s persistence in environmental conditions would need to be determined. Collaboration with Environment Canada and Health Canada would likely be required to introduce this novel bio-tag into current industrial processes.</li>
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<li></li>
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<li>When running test and dealing with proteins and Bacillus subtilis, all lab safety protocol needs to be followed at all times. This includes wearing the proper attire such as long pants, closed toe shoes, lab coats, lab gloves, and safety goggles or glasses. Once finished, all waste needs to be chemically and heat treated by bleaching and putting equipment, like beakers, into the autoclave. Other equipment, pipette tips, need to be disposed of correctly in biohazard bins.</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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<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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Revision as of 05:26, 17 October 2018

SAFETY

Precautions

The OLS igem Team has taken many precautions to ensure a safe lab environment, and the environmental safety of our new constructs. Some of the steps that have been taken to further ensure safety are:


  • In the design of the construct our team chose to use a lab strain of Bacillus subtilis, as it is a naturally occurring bacteria found in dirt and plants, so it is not harmful to the environment.
  • Our construct is intended for use as a protein bio-tag manufacturing system. The bio-tag protein would be isolated and purified for use, ensuring no live bacteria enter a sorting or recycling facility at any time.
  • In the process of applying our bio tag in the real world, the plastics will be bathed, rather than sprayed with the protein, to avoid airborne proteins and potential contamination.
  • Once plastic has been bio-tagged and sorted, it can now get recycled. In the recycling process plastic is shredded and melted down. The heat that is needed to melt the plastic will denature the proteins to stop potential environmental contamination. With future trials, the protein’s persistence in environmental conditions would need to be determined. Collaboration with Environment Canada and Health Canada would likely be required to introduce this novel bio-tag into current industrial processes.
  • When running test and dealing with proteins and Bacillus subtilis, all lab safety protocol needs to be followed at all times. This includes wearing the proper attire such as long pants, closed toe shoes, lab coats, lab gloves, and safety goggles or glasses. Once finished, all waste needs to be chemically and heat treated by bleaching and putting equipment, like beakers, into the autoclave. Other equipment, pipette tips, need to be disposed of correctly in biohazard bins.
Implemented prototype, what could be seen in a sorting facility.