Difference between revisions of "Team:Paris Bettencourt/Description"

 
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    Assay
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      <a href="https://2016.igem.org/Team:Paris_Bettencourt/Project/Microbiology" title="Microbiology">
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    Microbiology
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  Enzyme
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    Binding
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    Indigo
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    Model
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<div id=definition>
 
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<h1 class=project>Project description</h1>
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<h2 class="red">Short Summary: What We Did</h2>
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Dry cleaning is a process used to clean delicate fabrics which cannot withstand conventional detergents, physical forces and temperatures inside a washing machine. Despite being useful, dry cleaning can pose a threat to human health and the environment, since throughout the process different hazardous solvents are used to remove the stains from the fabric.
+
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Among the most widely used chemicals are tetrachloroethylene and perchloroethylene (also called "perc" by the industry). “Perc” is a volatile organic compound, hence it generates fumes that allow it to spread through the air from the clothes after the dry-cleaning process. Furthermore, “perc” is listed as "reasonably anticipated to be a human carcinogen" in the Thirteenth Report on Carcinogens published by the National Toxicology Program, because long-term exposure to perchloroethylene has been linked to different types of cancer. Luckily, the French government has begun the process of banning the use of “perc”, and promoting eco-friendly alternatives in all establishments close to the inhabited areas. <br>
+
  
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This is where our iGEM project starts. We interviewed several dry cleaning facilities in Paris to get a better insight about their needs, and what problems they might be facing. Not surprisingly, they told us that wine stains are one of the most difficult stains to remove from clothes, and thus we decided to have them as our main focus. <br>
+
<br>In this project we <a href="https://2016.igem.org/Team:Paris_Bettencourt/Project/Enzyme">produced synthetic enzymes</a> to remove red wine stains from fabric. These enzymes are designed to replace perchloroethylene (PERC), a toxic solvent used in dry cleaning that will soon be banned in France.<br>
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Wine, in general, is a complex mixture of alcohol, sugars, water and different secondary metabolites, such as anthocyanins, flavonoids, tannins, amongst many more. Our project is based on finding and improving enzymes, and processes, to degrade pigment molecules from wine stains. We are going to design and implement a series of assays in order to identify different enzymes and microbes from environmental samples. In addition, our project will develop a library of various binding domains with affinity for different fabrics, such as cotton, wool, silk, nylon or polyester. Some of these domains already exist in nature, and some are available in the iGEM registry. By adding these domains to our enzymes, we expect to improve cleaning efficiency and localization to the stain. We are also planning to focus on other important issues concerning our project, such as enzyme robustness, extraction and secretion, preserving the fabric quality, growth conditions and medias. Also, we will explore other interesting aspects of clothes industries like denim bleaching. And, of course, every successfully developed product will be forever perpetuated in the BioBrick format. <br>
+
 
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This project was selected from a pool of ideas generated in a series of discussions, brainstorming sessions and votings. Our selection process has included constant deducting and adding new ideas, up until final voting which resulted in the Frank‘n’Stain project. To achieve the best results on this year's iGEM competition, we have assembled a team based on our motivation, ambition, interdisciplinarity, creativity and scientific background. Together, with a little help from our advisors: Ariel Lindner, Jake Wintermute, Jason Bland and Nadine Bongaerts, we are going to work hard and rid the world of the nasty (although very French) wine stains! <br>
+
<br><a href="https://2016.igem.org/Team:Paris_Bettencourt/Integrated_Practices">We listened to people</a> who will use our product, conducting face-to-face interviews with every single dry cleaner in Paris. From this came a plan for a realistic product, a stain fighting enzymatic pretreatment compatible with existing cleaning technologies and workflows.<br>
−
Sources:<br>
+
 
−
  <a href="https://toxtown.nlm.nih.gov/text_version/chemicals.php?id=22">https://toxtown.nlm.nih.gov/text_version/chemicals.php?id=22 </a> <br>
+
<br><a href="https://2016.igem.org/Team:Paris_Bettencourt/Project/Microbiology">We isolated microbes</a> capable of degrading red wine from a library of 186 strains taken from vineyard soil samples collected around the world with the help of our fellow iGEMers. The most effective microbes were submitted for whole genome resequencing, then analyzed to produce a short list of candidate stain-fighting enzymes.<br>
 +
 
 +
<br><a href = "https://2016.igem.org/Team:Paris_Bettencourt/Model">We modeled enzymatic activity at the fabric surface</a> and determined that activity could be substantially improved if the enzymes had a moderate binding affinity for the fabric itself. This effectively increases the enzyme concentration at the fabric surface and reduces the quantity of enzymes lost through diffusion into the medium.<br>
 +
 
 +
<br><a href = "https://2016.igem.org/Team:Paris_Bettencourt/Project/Binding">We identified short peptides</a> with affinity to cotton, linen, wool, polyester and silk using the method of phage display. The resulting Fabric Binding Domains (FBDs) were quantitatively characterized using ELISA, to determine peptides with optimal affinity as determined by our model.<br>
 +
 
 +
<br><a href = "https://2016.igem.org/Team:Paris_Bettencourt/Project/Assay">We developed a high-throughput assay</a> to quantify stain removal on real cloth. Laser-cut fabric samples are sealed to 96-well microplates and imaged on flat-bed scanners. We coded custom image analysis software to identify each circular fabric sample and measure the stain intensity. <br>
 +
 
 +
<br><a href = "https://2016.igem.org/Team:Paris_Bettencourt/Parts">We constructed new BioBricks</a>, fusions of our most promising fabric binding domains to our favorite wine-degrading enzymes. The resulting proteins were expressed, purified and characterized both in vitro (in solution) and in situ (on real stained fabric).<br>
 +
 
 +
<br> We hope these pages convince you that our project is mature, thoroughly documented and effective. We believe that product is ready solve one small problem that real people have every day. <br>
  
−
  <a href="http://ntp.niehs.nih.gov/ntp/roc/content/listed_substances_508.pdf">http://ntp.niehs.nih.gov/ntp/roc/content/listed_substances_508.pdf</a> <br>
 
  
−
  <a href="http://www.developpement-durable.gouv.fr/Le-perchloroethylene-interdit-dans.html">http://www.developpement-durable.gouv.fr/Le-perchloroethylene-interdit-dans.html</a> <br>
 
 
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Latest revision as of 10:59, 1 December 2016


Short Summary: What We Did


In this project we produced synthetic enzymes to remove red wine stains from fabric. These enzymes are designed to replace perchloroethylene (PERC), a toxic solvent used in dry cleaning that will soon be banned in France.

We listened to people who will use our product, conducting face-to-face interviews with every single dry cleaner in Paris. From this came a plan for a realistic product, a stain fighting enzymatic pretreatment compatible with existing cleaning technologies and workflows.

We isolated microbes capable of degrading red wine from a library of 186 strains taken from vineyard soil samples collected around the world with the help of our fellow iGEMers. The most effective microbes were submitted for whole genome resequencing, then analyzed to produce a short list of candidate stain-fighting enzymes.

We modeled enzymatic activity at the fabric surface and determined that activity could be substantially improved if the enzymes had a moderate binding affinity for the fabric itself. This effectively increases the enzyme concentration at the fabric surface and reduces the quantity of enzymes lost through diffusion into the medium.

We identified short peptides with affinity to cotton, linen, wool, polyester and silk using the method of phage display. The resulting Fabric Binding Domains (FBDs) were quantitatively characterized using ELISA, to determine peptides with optimal affinity as determined by our model.

We developed a high-throughput assay to quantify stain removal on real cloth. Laser-cut fabric samples are sealed to 96-well microplates and imaged on flat-bed scanners. We coded custom image analysis software to identify each circular fabric sample and measure the stain intensity.

We constructed new BioBricks, fusions of our most promising fabric binding domains to our favorite wine-degrading enzymes. The resulting proteins were expressed, purified and characterized both in vitro (in solution) and in situ (on real stained fabric).

We hope these pages convince you that our project is mature, thoroughly documented and effective. We believe that product is ready solve one small problem that real people have every day.

Centre for Research and Interdisciplinarity (CRI)
Faculty of Medicine Cochin Port-Royal, South wing, 2nd floor
Paris Descartes University
24, rue du Faubourg Saint Jacques
75014 Paris, France
+33 1 44 41 25 22/25
igem2016parisbettencourt@gmail.com
2016.igem.org