Difference between revisions of "Team:TU Darmstadt"

 
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        <title> iGEM TU Darmstadt </title>
 
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        <div id="gemLO"><img src="https://static.igem.org/mediawiki/2017/9/9c/IGEM_logo_2500x2500.png" width="10%"></div>
 
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        <center><img src="https://static.igem.org/mediawiki/2017/d/dc/TUDarmstadtPDheader.png" width="50%"  ></center>
 
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        <h4>Chitosan is a biopolymer with both antibacterial and wound-healing properties. By linking fluorophores to chitosan oligomeres smart plasters can be produced, able to detect pathogenic bacteria via proteolytic activity. Therefore, production of designed chitosan for medical purpose is of special interest.
 
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        </h4><br>
 
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            <img src="https://static.igem.org/mediawiki/2017/3/36/TUDarmstadtPDplaster.png" alt="chitosan hydrogel" width="100%">
 
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        <h3 id="red">Engineering <i>E.&nbsp;coli</i> for specific synthesis of designer chitosan
 
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            Our primary wet-lab goal is to engineer a synthetic biological circuit for the specific synthesis of chitosans in <i>E.&nbsp;coli</i>. The chemical properties as well as the bioactivity of chitosans mainly depend on three variables:  the length of the oligomers, their level of deacetylation and their patterns of deacetylation. 
 
  
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<span class="image avatar"><a href="https://2017.igem.org/Team:TU_Darmstadt"><img src="https://static.igem.org/mediawiki/2017/3/3d/LogoOWL.png" alt="home"></a></span>
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<h1 id="logo"><a href="https://2017.igem.org/Team:TU_Darmstadt">ChiTUcare</a></h1>
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<li><a href="https://2017.igem.org/Team:TU_Darmstadt/project">Project</a></li>
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<li><a href="https://2017.igem.org/Team:TU_Darmstadt/human_practices">Human Practices</a></li>
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<li><a href="https://2017.igem.org/Team:TU_Darmstadt/tech">Tech</a></li>
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<li><a href="https://2017.igem.org/Team:TU_Darmstadt/team">Team</a></li>
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<li><a href="https://2017.igem.org/Team:TU_Darmstadt/judging">Judging</a></li>
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            Our enzymatic approach includes three enzymes. A chitin synthase (<i>Rhizobium&nbsp;leguminosarum bv.&nbsp;viciae</i>) catalyzes the oligomerization of N-acetylglucosamine-UDP monomers to chitin oligomers (tetramers and pentamers). 
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            Furthermore, two chitin deacetylases that differ in their regioselectivity (<i>Sinorhizobium&nbsp;meliloti</i>  (nodB) and <i>Puccina&nbsp;graminis f.&nbsp;sp.&nbsp;tritici</i>) are regulated orthogonally, making it possible to choose between two different types of deacetylation patterns.
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            <img src="https://static.igem.org/mediawiki/2017/2/2d/TUDarmstadtPDchitosansynthesis.png" alt="pathway chitosan" width="100%">
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         <h3 id="red">Application of chitosan oligomers:
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<img src="https://static.igem.org/mediawiki/2017/6/63/T--TU_Darmstadt--BannerHomePage5.png" style="width:100%;height:auto;"/>
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            chitosan hydrogel for bacterial enzyme detection
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            Downstream of the synthetic biological circuit for the synthesis of chitosan pentamers, we want to give an example for an explicit application of chitosan oligomers. This shall be accomplished by building a plaster for wounds carrying the chitosan hydrogel with chitosan oligomers linked to a fluorophore via a peptide chain. When the plaster is applied to a wound it can detect bacterial protease activities and thus diagnose wound infection. Proteases will cleave the peptide linker and release the fluorophore, that is then detectable via UV-light.
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  <center> <a href="https://2017.igem.org/Team:TU_Darmstadt/Demonstrate">
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  <h2 class="HoverText"> Proof of Concept </h2></a> </center>
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<div class="4u"><span class="image fit"><a href="https://2017.igem.org/Team:TU_Darmstadt/project/hydrogel"><img src="https://static.igem.org/mediawiki/2017/e/e3/T--TU_Darmstadt--Teaser_Pic4.png" style="border-radius: 50%;" class="HoverBorder" alt=""/></a></span></div>
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                <div class="6u"><span class="image fit"><video src="https://static.igem.org/mediawiki/2017/5/56/T--TU_Darmstadt--geiles_video.mp4" alt="Chitosan-Alkaline production" style="
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<div class="4u"><span class="image fit"><a href="https://2017.igem.org/Team:TU_Darmstadt/project/chitin_deacetylase"><img src="https://static.igem.org/mediawiki/2017/c/cf/T--TU_Darmstadt--Teaser_Pic2.png" style="border-radius: 50%;"  class="HoverBorder" alt="" /></a></span></div>
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<div class="4u"><span class="image fit"><a href="https://2017.igem.org/Team:TU_Darmstadt/project/chemistry"><img src="https://static.igem.org/mediawiki/2017/d/db/T--TU_Darmstadt--Teaser_Pic1.png" style="border-radius: 50%;"  class="HoverBorder" alt="" /></a></span></div>
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<p>Chitosan is a biopolymer with both antibacterial and wound-healing properties. By linking fluorophores to chitosan oligomeres smart plasters can be produced, able to detect pathogenic bacteria via proteolytic activity. Therefore, production of designed chitosan for medical purpose is of special interest.</p>
 +
<center><img src="https://static.igem.org/mediawiki/2017/3/36/TUDarmstadtPDplaster.png" alt="chitosan hydrogel" width="80%" style="padding: 1em 0 1em 0;"></center>
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<h4>Engineering <i>E.&nbsp;coli</i> for specific Synthesis of Designer Chitosan</h4>
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<p>Our primary wet-lab goal is to engineer a synthetic biological circuit for the specific synthesis of chitosans in <i>E.&nbsp;coli</i>. The chemical properties as well as the bioactivity of chitosans mainly depend on three variables:  the length of the oligomers, their level of deacetylation and their patterns of deacetylation.</p>
 +
        <p>Our enzymatic approach includes three enzymes. A chitin synthase (<i>Rhizobium&nbsp;leguminosarum bv.&nbsp;viciae</i>) catalyzes the oligomerization of N-acetylglucosamine-UDP monomers to chitin oligomers (tetramers and pentamers). Furthermore, two chitin deacetylases that differ in their regioselectivity (<i>Sinorhizobium&nbsp;meliloti</i>  (nodB) and <i>Puccina&nbsp;graminis f.&nbsp;sp.&nbsp;tritici</i>) are regulated orthogonally, making it possible to choose between two different types of deacetylation patterns.</p>
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            <center><img src="https://static.igem.org/mediawiki/2017/2/2d/TUDarmstadtPDchitosansynthesis.png" alt="pathway chitosan" width="80%" style="padding: 1em 0 1em 0;"></center>
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<h4>Application of Chitosan Oligomers: Chitosan Hydrogel for bacterial Enzyme Detection</h4>
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        <p>Downstream of the synthetic biological circuit for the synthesis of chitosan pentamers, we want to give an example for an explicit application of chitosan oligomers. This shall be accomplished by building a plaster for wounds carrying the chitosan hydrogel with chitosan oligomers linked to a fluorophore via a peptide chain. When the plaster is applied to a wound it can detect bacterial protease activities and thus diagnose wound infection. Proteases will cleave the peptide linker and release the fluorophore, that is then detectable via UV-light.</p>
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            <center><img src="https://static.igem.org/mediawiki/2017/4/40/TUDarmstadtPDpeptidelinker.png" alt="peptide linker" width="50%" style="padding: 1em 0 1em 0;"></center>
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Latest revision as of 20:27, 1 November 2017

MainPage

image/svg+xml 1. Chitosan:Chitosan is a derivate of the linear polysaccharide chitin. The chemical and physical properties can vary remarkably. This enables a huge scope of applications. Chitosan shows antimicrobial properties and supports scar free wound healing.Visit the subpage! 2. Wound infection:Simple and cheap wound treatment is a challenging task.The detection of potentially present pathogenic bacteria iscomplicated, as removal of the wound dressing disrupts thehealing process. 3. Chitin Synthase:To realize the synthesis of chitosan, for usage in wounddressing hydrogels, we produce chitin out of N-acetyl-glucosamine-UDP monomers. Chitin is a precursor for chitosan.We implement the chitin synthase NodC, which createstetrameric and pentameric chitin.Visit the subpage! 4. Chitin Deacetylase:To manufacture chitosan out of chitin, the producedchitin oligomers have to be deacetylated. This can beperformed by using certain hydrolyzing enzymes called chitin deacetylases. In our case,we use NodB and COD.Visit the subpage! 5. Chemistry:To deal with the problem of wound infections andtheir detection, our synthesized chitosan is linkedto a flourophor via a peptide linker. This makes itpossible to detect exoproteases from pathogenicbacteria within minutes.Visit the subpage! image/svg+xml 6. Hydrogel:To use the wound healing supportive properties of chitosan,we manufactured non-toxic, low-cost hydrogels, containingdefined chitosans for usage in wound care.Visit the subpage! 9. CloneCademy:In order to share the knowledge aboutsynthetic biology and the achievementsof our project, we developed a web-basedinteractive learning platform calledCloneCademy. This education tool makesit possible for other iGEM teams to sharetheir ideas with society.Visit the subpage! 7. Solution:By combining the physiological propertiesof chitosan and the novel detectionsystem for wound infections in a hydrogelbandaid, we realized next generation wound care. Furthermore, we successfullyproved all milestones of our project.Visit the subpage! 8. Tech:We constructed a smartphone-adaptable, low-cost and mainly 3Dprinted microscope with a µm resolutionby implementing the digital inlineholographic approach. Thus, allowing the analyzation of ourhydrogel structure during the project. We also presenta software solution enabling 3D analyzation based onthe open source HoloPy package.Visit the subpage!


Proof of Concept