Team:Wageningen UR/Summary

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<div class="projectheader">
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<img src="https://static.igem.org/mediawiki/2013/1/13/Aspergillusheader.png">
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<div class="content summary">
<div class="content summary">
<h1>Summary</h1>
<h1>Summary</h1>
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<h2>Subtitle</h2>
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<h2> <i>We made mistakes, <br/>though this whole endeavor was incredibly worthwhile.<br/>
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We learned a lot,<br/> and nothing beats the excitement of positive results.</i><br/><br/> -- Wageningen UR 2013 iGEM students</h2>
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<div class="tournav">
<div class="tournav">
     <ul>
     <ul>
-
         <li class="first bgsm fill"><a href="https://2013.igem.org/Team:Wageningen_UR/Why_Aspergillus_nigem">Why <i>Aspergillus nigem</i>?</a></li>
+
         <li class="firstbgsm fill"><a href="https://2013.igem.org/Team:Wageningen_UR/Why_Aspergillus_nigem">Why <i>Aspergillus nigem</i>?</a></li>
-
         <li class="fill">Secondary metabolites</li>
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         <li class="fill"><a href="https://2013.igem.org/Team:Wageningen_UR/Secondary_metabolites">Secondary metabolites</a></li>
-
         <li class="fill">Lovastatin</li>
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         <li class="fill"><a href="https://2013.igem.org/Team:Wageningen_UR/Lovastatin">Lovastatin</a></li>
-
         <li class="fill">ATP Biosensor</li>
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         <li class="fill"><a href="https://2013.igem.org/Team:Wageningen_UR/Modeling">Modeling</a></li>
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         <li class="fill">pH Biosensor</li>
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         <li class="fill"><a href="https://2013.igem.org/Team:Wageningen_UR/Biosensors">Biosensors</a></li>
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         <li class="fill">Cytoskeleton and Septa</li>
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         <li class="fill"><a href="https://2013.igem.org/Team:Wageningen_UR/Infrastructure">Infrastructure</a></li>
-
         <li class="fill">Chromoproteins</li>
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         <li class="fill"><a href="https://2013.igem.org/Team:Wageningen_UR/Chromoproteins">Chromoproteins</a></li>
-
         <li class="fill">Host engineering</li>
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         <li class="fill"><a href="https://2013.igem.org/Team:Wageningen_UR/Host_engineering">Host engineering</a></li>
         <li class="last current">Summary</li>
         <li class="last current">Summary</li>
     </ul>
     </ul>
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<div class="tournav">
<div class="tournav">
     <ul>
     <ul>
-
         <li class="first fill">Why <i>Aspergillus nigem</i>?</li>
+
         <li class="first fill"><a href="https://2013.igem.org/Team:Wageningen_UR/Why_Aspergillus_nigem">Why <i>Aspergillus nigem</i>?</a></li>
         <li class="smbg fill"><a href="https://2013.igem.org/Team:Wageningen_UR/Secondary_metabolites">Secondary metabolites</a></li>
         <li class="smbg fill"><a href="https://2013.igem.org/Team:Wageningen_UR/Secondary_metabolites">Secondary metabolites</a></li>
         <li class="bgbg fill"><a href="https://2013.igem.org/Team:Wageningen_UR/Lovastatin">Lovastatin</a></li>
         <li class="bgbg fill"><a href="https://2013.igem.org/Team:Wageningen_UR/Lovastatin">Lovastatin</a></li>
-
         <li class="bgsm fill"><a href="https://2013.igem.org/Team:Wageningen_UR/ATP_biosensor">ATP Biosensor</a></li>
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         <li class="bgsm fill"><a href="https://2013.igem.org/Team:Wageningen_UR/Modeling">Modeling</a></li>
-
         <li class="fill">pH Biosensor</li>
+
         <li class="fill"><a href="https://2013.igem.org/Team:Wageningen_UR/Biosensors">Biosensors</a></li>
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         <li class="fill">Cytoskeleton and Septa</li>
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         <li class="fill"><a href="https://2013.igem.org/Team:Wageningen_UR/Infrastructure">Infrastructure</a></li>
-
         <li class="fill">Chromoproteins</li>
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         <li class="fill"><a href="https://2013.igem.org/Team:Wageningen_UR/Chromoproteins">Chromoproteins</a></li>
-
         <li class="fill">Host engineering</li>
+
         <li class="fill"><a href="https://2013.igem.org/Team:Wageningen_UR/Host_engineering">Host engineering</a></li>
         <li class="last current">Summary</li>
         <li class="last current">Summary</li>
     </ul>
     </ul>
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<div class="tournav">
<div class="tournav">
     <ul>
     <ul>
-
         <li class="first fill">Why <i>Aspergillus nigem</i>?</li>
+
         <li class="first fill"><a href="https://2013.igem.org/Team:Wageningen_UR/Why_Aspergillus_nigem">Why <i>Aspergillus nigem</i>?</a></li>
-
         <li class="fill">Secondary metabolites</li>
+
         <li class="fill"><a href="https://2013.igem.org/Team:Wageningen_UR/Secondary_metabolites">Secondary metabolites</a></li>
-
         <li class="fill">Lovastatin</li>
+
         <li class="fill"><a href="https://2013.igem.org/Team:Wageningen_UR/Lovastatin">Lovastatin</a></li>
-
         <li class="fill">ATP Biosensor</li>
+
         <li class="fill"><a href="https://2013.igem.org/Team:Wageningen_UR/Modeling">Modeling</a></li>
-
         <li class="smbg fill"><a href="https://2013.igem.org/Team:Wageningen_UR/pH_biosensor">pH Biosensor</a></li>
+
         <li class="smbg fill"><a href="https://2013.igem.org/Team:Wageningen_UR/Biosensors">Biosensors</a></li>
-
         <li class="bgbg fill"><a href="https://2013.igem.org/Team:Wageningen_UR/Cytoskeleton_and_septa">Cytoskeleton and Septa</a></li>
+
         <li class="bgbg fill"><a href="https://2013.igem.org/Team:Wageningen_UR/Infrastructure">Infrastructure</a></li>
         <li class="bgsm fill"><a href="https://2013.igem.org/Team:Wageningen_UR/Chromoproteins">Chromoproteins</a></li>
         <li class="bgsm fill"><a href="https://2013.igem.org/Team:Wageningen_UR/Chromoproteins">Chromoproteins</a></li>
-
         <li class="fill">Host engineering</li>
+
         <li class="fill"><a href="https://2013.igem.org/Team:Wageningen_UR/Host_engineering">Host engineering</a></li>
         <li class="last current">Summary</li>
         <li class="last current">Summary</li>
     </ul>
     </ul>
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<div class="tournav">
<div class="tournav">
     <ul>
     <ul>
-
         <li class="first fill">Why <i>Aspergillus nigem</i>?</li>
+
         <li class="first fill"><a href="https://2013.igem.org/Team:Wageningen_UR/Why_Aspergillus_nigem">Why <i>Aspergillus nigem</i>?</a></li>
-
         <li class="fill">Secondary metabolites</li>
+
         <li class="fill"><a href="https://2013.igem.org/Team:Wageningen_UR/Secondary_metabolites">Secondary metabolites</a></li>
-
         <li class="fill">Lovastatin</li>
+
         <li class="fill"><a href="https://2013.igem.org/Team:Wageningen_UR/Lovastatin">Lovastatin</a></li>
-
         <li class="fill">ATP Biosensor</li>
+
         <li class="fill"><a href="https://2013.igem.org/Team:Wageningen_UR/Modeling">Modeling</a></li>
-
         <li class="fill">pH Biosensor</li>
+
         <li class="fill"><a href="https://2013.igem.org/Team:Wageningen_UR/Biosensors">Biosensors</a></li>
-
         <li class="fill">Cytoskeleton and Septa</li>
+
         <li class="fill"><a href="https://2013.igem.org/Team:Wageningen_UR/Infrastructure">Infrastructure</a></li>
-
         <li class="fill">Chromoproteins</li>
+
         <li class="fill"><a href="https://2013.igem.org/Team:Wageningen_UR/Chromoproteins">Chromoproteins</a></li>
-
         <li class="smbgsm fill">Host engineering</li>
+
         <li class="smbgsm fill"><a href="https://2013.igem.org/Team:Wageningen_UR/Host_engineering">Host engineering</a></li>
         <li class="last current">Summary</li>
         <li class="last current">Summary</li>
     </ul>
     </ul>
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<div class="tournav">
<div class="tournav">
     <ul>
     <ul>
-
         <li class="first fill">Why <i>Aspergillus nigem</i>?</li>
+
         <li class="first fill"><a href="https://2013.igem.org/Team:Wageningen_UR/Why_Aspergillus_nigem">Why <i>Aspergillus nigem</i>?</a></li>
-
         <li class="fill">Secondary metabolites</li>
+
         <li class="fill"><a href="https://2013.igem.org/Team:Wageningen_UR/Secondary_metabolites">Secondary metabolites</a></li>
-
         <li class="fill">Lovastatin</li>
+
         <li class="fill"><a href="https://2013.igem.org/Team:Wageningen_UR/Lovastatin">Lovastatin</a></li>
-
         <li class="fill">ATP Biosensor</li>
+
         <li class="fill"><a href="https://2013.igem.org/Team:Wageningen_UR/Modeling">Modeling</a></li>
-
         <li class="fill">pH Biosensor</li>
+
         <li class="fill"><a href="https://2013.igem.org/Team:Wageningen_UR/Biosensors">Biosensors</a></li>
-
         <li class="fill">Cytoskeleton and Septa</li>
+
         <li class="fill"><a href="https://2013.igem.org/Team:Wageningen_UR/Infrastructure">Infrastructure</a></li>
-
         <li class="fill">Chromoproteins</li>
+
         <li class="fill"><a href="https://2013.igem.org/Team:Wageningen_UR/Chromoproteins">Chromoproteins</a></li>
-
         <li class="fill">Host engineering</li>
+
         <li class="fill"><a href="https://2013.igem.org/Team:Wageningen_UR/Host_engineering">Host engineering</a></li>
         <li class="last current">Summary</li>
         <li class="last current">Summary</li>
     </ul>
     </ul>
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<div id="pagewrapper">
<div id="pagewrapper">
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    <p>Concluding our iGEM project in 2013, we introduced <i>Aspergillus niger</i> as a potential host into iGEM. It is an industrially relevant organism, deserving a place in iGEM as a standard synthetic biology chassis. With the host engineering, we stepped further to the next level of synthetic biology.</p>
 +
 +
<p>In this GRAS organism, we established a modular system of domain shuffling in order to express a variety of secondary metabolites. We focused on the production of a medically relevant compound, lovastatin, which has never been produced in <i>A. niger</i>.We synthesized a range of modules for the main multi-domain enzyme in the lovastatin pathway. We also developed a new frameshiftless assembly protocol to design new multidomain enzyme complexes, and bricked these parts for future teams.<p>
 +
 +
<p>Additionally, to target the production in a specific compartment, a toolkit including a <a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1023001" target="_blank"> promoter</a>, <a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1023002" target="_blank"> terminator</a>, biosensors for <a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1023004" target="_blank"> pH</a>, <a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1023003 " target="_blank"> ATP</a>, and <a href="http://parts.igem.org/wiki/index.php?title=Part:BBa_K1023005" target="_blank"> chromogenic biomarkers</a>, as well as a marker for the <a href=" http://parts.igem.org/wiki/index.php?title=Part:BBa_K1023006" target="_blank"> cytoskeleton</a> was created. It paved the way for future teams to further exploit this beautiful bug. Furthermore, the metabolic models we developed to pre-emptively assess the production conditions, and potentially the optimization of production, has hinted at future improvements for <i>A. niger</i> lovastatin production.</p>
 +
 +
<p>Finally, we modified the host by directed evolution. Filamentous fungi are pretty organisms, but multicellularity is not always wanted in fermentations. We set out to investigate it by evolving mutants with a reduced mycelial cohesiveness. Microscopic analyses have shown that <i>Aspergillus</i> is conditionally dimorphic, and both viable and metabolically active in a single cell morphotype. We made important steps in unravelling the causes of multicellularity. </p>
 +
 +
<p>All in all, our project has been ambitious, inspiring, educational, social, and totally worthwhile. We made mistakes, underestimated some experimental time frames, totally failed in some experimental set ups, but nothing beats the excitement of positive results.<p> 
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Latest revision as of 03:43, 5 October 2013

Summary

We made mistakes,
though this whole endeavor was incredibly worthwhile.
We learned a lot,
and nothing beats the excitement of positive results.

-- Wageningen UR 2013 iGEM students

Concluding our iGEM project in 2013, we introduced Aspergillus niger as a potential host into iGEM. It is an industrially relevant organism, deserving a place in iGEM as a standard synthetic biology chassis. With the host engineering, we stepped further to the next level of synthetic biology.

In this GRAS organism, we established a modular system of domain shuffling in order to express a variety of secondary metabolites. We focused on the production of a medically relevant compound, lovastatin, which has never been produced in A. niger.We synthesized a range of modules for the main multi-domain enzyme in the lovastatin pathway. We also developed a new frameshiftless assembly protocol to design new multidomain enzyme complexes, and bricked these parts for future teams.

Additionally, to target the production in a specific compartment, a toolkit including a promoter, terminator, biosensors for pH, ATP, and chromogenic biomarkers, as well as a marker for the cytoskeleton was created. It paved the way for future teams to further exploit this beautiful bug. Furthermore, the metabolic models we developed to pre-emptively assess the production conditions, and potentially the optimization of production, has hinted at future improvements for A. niger lovastatin production.

Finally, we modified the host by directed evolution. Filamentous fungi are pretty organisms, but multicellularity is not always wanted in fermentations. We set out to investigate it by evolving mutants with a reduced mycelial cohesiveness. Microscopic analyses have shown that Aspergillus is conditionally dimorphic, and both viable and metabolically active in a single cell morphotype. We made important steps in unravelling the causes of multicellularity.

All in all, our project has been ambitious, inspiring, educational, social, and totally worthwhile. We made mistakes, underestimated some experimental time frames, totally failed in some experimental set ups, but nothing beats the excitement of positive results.

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