Difference between revisions of "Team:UMaryland/PETaseIntro"

 
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<div class="row"> <!-- honestly idk why this is here - i just copied it from somewhere and then now i'm too scared to remove this class !-->
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<div class="titleRegion" style="background-image: url(https://static.igem.org/mediawiki/2018/6/66/T--UMaryland--patternAlternateTwo.png)">
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<div class="container" style="height: 200px;">
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<div class="titleText">History</div>
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<div class="subtitleText">Past iGEM Teams involved with PET</div>
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<div id="Overview">
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<div class="meatMeat">
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Listed here are past iGEM teams that have worked on projects on or related to PET degredation.
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</div>
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</div>
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<div id="2012">
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<div class="meatSubtitle">
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2012
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</div>
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<div class="meatMeat">
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<a href="https://2012.igem.org/Team:BAU-Indonesia">BAU-Indonesia</a>
 +
<ul>
 +
<li>Isolation of cutinase gene from nature with primers</li>
 +
</ul><br>
 +
<a href="https://2012.igem.org/Team:TU_Darmstadt">TU Darmstadt</a>
 +
<ul>
 +
<li>Surface display of cutinase on E. coli</li>
 +
<li>Attempted TPA transport into E. coli, further research required</li>
 +
<li>Expressed all TPH enzymes, did not attempt to measure activity</li>
 +
<li>Confirmed anaerobic conversion of PCA via AroY and  XylE enzymes</li>
 +
</ul><br>
 +
<a href="https://2012.igem.org/Team:UC_Davis">UC Davis</a>
 +
<ul>
 +
<li>Confirmed cutinase activity using PNPB esterase assay</li>
 +
<li>Engineered E. coli ethylene glycol metabolism with directed evolution</li>
 +
</ul><br>
 +
</div>
 +
</div>
 +
<div id="2013">
 +
<div class="meatSubtitle">
 +
2013
 +
</div>
 +
<div class="meatMeat">
 +
<a href="https://2013.igem.org/Team:Imperial_College">Imperial College</a>
 +
<ul>
 +
<li>Produced P3HB bioplastic from mixed waste containing at least some PET</li>
 +
</ul><br>
 +
</div>
 +
</div>
 +
<div id="2014">
 +
<div class="meatSubtitle">
 +
2014
 +
</div>
 +
<div class="meatMeat">
 +
<a href="https://2014.igem.org/Team:METU_Turkey">METU Turkey</a>
 +
<ul>
 +
<li>Reduced catechol to pyruvate</li>
 +
</ul><br>
 +
<a href="https://2014.igem.org/Team:ITB_Indonesia">ITB Indonesia</a>
 +
<ul>
 +
<li>LC cutinatse activity confimed with SEM, PNPB</li>
 +
</ul><br>
 +
</div>
 +
</div>
 +
<div id="2015">
 +
<div class="meatSubtitle">
 +
2015
 +
</div>
 +
<div class="meatMeat">
 +
<a href="https://2015.igem.org/Team:Pasteur_Paris">Pasteur Paris</a>
 +
<ul>
 +
<li>PNPB assay to confirm activity of esterase EST13</li>
 +
<li>Fluorescent detection of TPA can not be accomplished when in LB broth</li>
 +
</ul><br>
 +
</div>
 +
</div>
 +
<div id="2016">
 +
<div class="meatSubtitle">
 +
2016
 +
</div>
 +
<div class="meatMeat">
 +
<a href="https://2016.igem.org/Team:ASIJ_Tokyo">ASIJ Tokyo</a>
 +
<ul>
 +
<li>Attempt at detecting PET degradation by mass change failed</li>
 +
</ul><br>
 +
<a href="https://2016.igem.org/Team:AUC_TURKEY">AUC Turkey</a>
 +
<ul>
 +
<li><i>Withdrawn</i></li>
 +
</ul><br>
 +
<a href="https://2016.igem.org/Team:Baltimore_BioCrew">Baltimore BioCrew</a>
 +
<ul>
 +
<li>Planned to weigh PET degradation, no results</li>
 +
</ul><br>
 +
<a href="https://2016.igem.org/Team:BGU_ISRAEL">BGU Israel</a>
 +
<ul>
 +
<li>PNPB and EM to confirm LC cutinase activity</li>
 +
<li>P. putida can grow on PCA as sole carbon source, but not TPA</li>
 +
<li>E. coli expressing LC-cutinase with pelB leader sequence grew on M9 plates with PET as sole carbon source. Expected to be due to consumption of ethylene glycol from PET degradation</li>
 +
<li>Unable to determine enzyme efficiency based on growth due to heterogeneity in PET distribution</li>
 +
<li>Measured fluorescence of TPA on plates, unable to quantify LC cutinase activity</li>
 +
</ul><br>
 +
<a href="https://2016.igem.org/Team:Harvard_BioDesign">Harvard BioDesign</a>
 +
<ul>
 +
<li>Petase function confirmed with PNPB</li>
 +
<li>Bacteria produced electric current when supplied with unspecified quantity of TPA</li>
 +
</ul><br>
 +
<a href="https://2016.igem.org/Team:Tianjin">Tianjin</a>
 +
<ul>
 +
<li>EM confirmation of PETase activity of PET film degradation</li>
 +
<li>Multispectral scanning quantified PETase products for cell free system</li>
 +
</ul><br>
 +
<a href="https://2016.igem.org/Team:TJUSLS_China">TJUSLS China</a>
 +
<ul>
 +
<li>HPLC detection of MHET to confirm PETase activity in varying conditions</li>
 +
<li>Surface display of PETase in E. coli</li>
 +
</ul><br>
 +
<a href="https://2016.igem.org/Team:UESTC-China">UESTC-China</a>
 +
<ul>
 +
<li>SEM and PNPB to confirm PETase activity</li>
 +
<li>Possible detection of TPA by UV vis (higher absorbance across spectrum)</li>
 +
</ul><br>
 +
<a href="https://2016.igem.org/Team:UoA_NewZealand">UoA New Zealand</a>
 +
<ul>
 +
<li>Assembled PETase part with His tag</li>
 +
</ul><br>
 +
</div>
 +
</div>
 +
<div id="2017">
 +
<div class="meatSubtitle">
 +
2017
 +
</div>
 +
<div class="meatMeat">
 +
<a href="https://2017.igem.org/Team:Baltimore_Bio-Crew">Baltimore Bio-Crew</a>
 +
<ul>
 +
<li>Fluorescine diacetate hydrolysis assay to confirm PETase and MHETase hydrolytic activity</li>
 +
</ul><br>
 +
<a href="https://2017.igem.org/Team:BOKU-Vienna">BOKU-Vienna</a>
 +
<ul>Discussion of a possible method for directed evolution of PETase by culturing cells on PET film that fluoresces when degraded</li>
 +
</ul><br>
 +
<a href="https://2017.igem.org/Team:ITB_Indonesia">ITB Indonesia</a>
 +
<ul>
 +
<li>Successful biofilm formation on PET, but biofilm matrix hampered PETase activity</li>
 +
<li>Engineered E. coli ethylene glycol metabolism with directed evolution</li>
 +
</ul><br>
 +
</div>
 +
</div>
 +
<div id="2018">
 +
<div class="meatSubtitle">
 +
2018
 +
</div>
 +
<div class="meatMeat">
 +
<a href="https://2018.igem.org/Team:Makerere_University">Makerere University</a>
 +
<ul>
 +
<li><i>TBA</i></li>
 +
</ul><br>
 +
<a href="https://2018.igem.org/Team:OLS_Canmore_Canada">OLS Canmore Canada</a>
 +
<ul>
 +
<li><i>TBA</i></li>
 +
</ul><br>
 +
<a href="https://2018.igem.org/Team:RHIT">RHIT</a>
 +
<ul>
 +
<li><i>TBA</i></li>
 +
</ul><br>
 +
<a href="https://2018.igem.org/Team:ULaVerne_Collab">ULaVerne</a>
 +
<ul>
 +
<li><i>TBA</i></li>
 +
</ul><br>
 +
<a href="https://2018.igem.org/Team:UMaryland">UMaryland</a>
 +
<ul>
 +
<li>Check out our results on this wiki!</li>
 +
</ul><br>
 +
<a href="https://2018.igem.org/Team:Yale">Yale</a>
 +
<ul>
 +
<li><i>TBA</i></li>
 +
</ul><br>
 +
</div>
 +
</div>
 +
</div>
 +
</div>
 +
</div>
 +
</body>
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<div class="column full_size">
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<h1>History</h1>
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<h3>iGEM teams pursuing PET related projects</h3>
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<img src="https://static.igem.org/mediawiki/2018/8/88/T--UMaryland--PETasehist.png">
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<p>
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SUMMARY OF WET LAB RESULTS
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−
<p>
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−
<p>
+
−
UC DAVIS 2012
+
−
<p>
+
−
<p>
+
−
        Confirmed cutinase activity using PNPB esterase assay
+
−
<p>
+
−
<p>
+
−
        Engineered E. coli ethylene glycol metabolism with directed evolution
+
−
<p>
+
−
<p>
+
−
BAU-Indonesia 2012
+
−
<p>
+
−
<p>
+
−
        Isolation of cutinase gene from nature with primers
+
−
<p>
+
−
<p>
+
−
TU Darmstadt 2012
+
−
<p>
+
−
<p>
+
−
Surface display of cutinase on E. coli
+
−
<p>
+
−
<p>
+
−
Attempted TPA transport into E. coli, further research required
+
−
<p>
+
−
<p>
+
−
Expressed all TPH enzymes, did not attempt to measure activity
+
−
<p>
+
−
<p>
+
−
Confirmed anaerobic conversion of PCA via AroY and  XylE enzymes
+
−
<p>
+
−
<p>
+
−
Imperial_College 2013
+
−
<p>
+
−
<p>
+
−
Produced P3HB bioplastic from mixed waste containing at least some PET
+
−
<p>
+
−
<p>
+
−
METU_Turkey 2014
+
−
<p>
+
−
<p>
+
−
Reduced catechol to pyruvate
+
−
<p>
+
−
<p>
+
−
ITB-Indonesia 2014
+
−
<p>
+
−
<p>
+
−
LC cutinatse activity confimed with SEM, PNPB
+
−
<p>
+
−
<p>
+
−
Pasteur Paris 2015
+
−
<p>
+
−
<p>
+
−
PNPB assay to confirm activity of esterase EST13
+
−
<p>
+
−
<p>
+
−
Fluorescent detection of TPA can not be accomplished when in LB broth.
+
−
<p>
+
−
<p>
+
−
Harvard 2016
+
−
<p>
+
−
<p>
+
−
Petase function confirmed with PNPB
+
−
<p>
+
−
<p>
+
−
Bacteria produced electric current when supplied with unspecified quantity of TPA
+
−
<p>
+
−
<p>
+
−
ASIJ Tokyo 2016
+
−
<p>
+
−
<p>
+
−
Attempt at detecting PET degradation by mass change failed
+
−
<p>
+
−
<p>
+
−
UoA_NewZealand 2016
+
−
<p>
+
−
<p>
+
−
Assembled PETase part with His tag
+
−
<p>
+
−
<p>
+
−
BGU-Israel 2016
+
−
<p>
+
−
<p>
+
−
PNPB and EM to confirm LC cutinase activity
+
−
<p>
+
−
<p>
+
−
P. putida can grow on PCA as sole carbon source, but not TPA.
+
−
<p>
+
−
<p>
+
−
E. coli expressing LC-cutinase with pelB leader sequence grew on M9 plates with PET as sole carbon source. Expected to be due to consumption of ethylene glycol from PET degradation.
+
−
<p>
+
−
<p>
+
−
Unable to determine enzyme efficiency based on growth due to heterogeneity in PET distribution
+
−
<p>
+
−
<p>
+
−
Measured fluorescence of TPA on plates, unable to quantify LC cutinase activity.
+
−
<p>
+
−
<p>
+
−
TJUSLS China 2016
+
−
<p>
+
−
<p>
+
−
HPLC detection of MHET to confirm PETase activity in varying conditions
+
−
<p>
+
−
<p>
+
−
Surface display of PETase in E. coli
+
−
<p>
+
−
<p>
+
−
Tianjin 2016
+
−
<p>
+
−
<p>
+
−
EM confirmation of PETase activity of PET film degradation
+
−
<p>
+
−
<p>
+
−
Multispectral scanning quantified PETase products for cell free system
+
−
<p>
+
−
<p>
+
−
Baltimore BioCrew 2016
+
−
<p>
+
−
<p>
+
−
Planned to weigh PET degradation, no results
+
−
<p>
+
−
<p>
+
−
UESTC China 2016
+
−
<p>
+
−
<p>
+
−
SEM and PNPB to confirm PETase activity
+
−
<p>
+
−
<p>
+
−
Possible detection of TPA by UV vis (higher absorbance across spectrum)
+
−
<p>
+
−
<p>
+
−
AUC_Turkey 2016
+
−
<p>
+
−
<p>
+
−
Withdrawn
+
−
<p>
+
−
<p>
+
−
ITB-Indonesia 2017
+
−
<p>
+
−
<p>
+
−
PNPB and SEM to confirm PETase activity
+
−
<p>
+
−
<p>
+
−
Successful biofilm formation on PET, but biofilm matrix hampered PETase activity.
+
−
<p>
+
−
<p>
+
−
Baltimore BioCrew 2017
+
−
<p>
+
−
<p>
+
−
Fluorescine diacetate hydrolysis assay to confirm PETase and MHETase hydrolytic activity
+
−
<p>
+
−
<p>
+
−
BOKU-Vienna 2017
+
−
<p>
+
−
<p>
+
−
Discussion of a possible method for directed evolution of PETase by culturing cells on PET film that fluoresces when degraded
+
−
<p>
+
−
<p>
+
−
If you see this then 2018 results from other teams have not been posted in time to show them here before the wiki freeze.
+
−
 
+
−
<p>
+

Latest revision as of 22:45, 15 October 2018

Template Title Template Title

History
Past iGEM Teams involved with PET
Listed here are past iGEM teams that have worked on projects on or related to PET degredation.
2012
BAU-Indonesia
  • Isolation of cutinase gene from nature with primers

TU Darmstadt
  • Surface display of cutinase on E. coli
  • Attempted TPA transport into E. coli, further research required
  • Expressed all TPH enzymes, did not attempt to measure activity
  • Confirmed anaerobic conversion of PCA via AroY and XylE enzymes

UC Davis
  • Confirmed cutinase activity using PNPB esterase assay
  • Engineered E. coli ethylene glycol metabolism with directed evolution

2013
Imperial College
  • Produced P3HB bioplastic from mixed waste containing at least some PET

2014
METU Turkey
  • Reduced catechol to pyruvate

ITB Indonesia
  • LC cutinatse activity confimed with SEM, PNPB

2015
Pasteur Paris
  • PNPB assay to confirm activity of esterase EST13
  • Fluorescent detection of TPA can not be accomplished when in LB broth

2016
ASIJ Tokyo
  • Attempt at detecting PET degradation by mass change failed

AUC Turkey
  • Withdrawn

Baltimore BioCrew
  • Planned to weigh PET degradation, no results

BGU Israel
  • PNPB and EM to confirm LC cutinase activity
  • P. putida can grow on PCA as sole carbon source, but not TPA
  • E. coli expressing LC-cutinase with pelB leader sequence grew on M9 plates with PET as sole carbon source. Expected to be due to consumption of ethylene glycol from PET degradation
  • Unable to determine enzyme efficiency based on growth due to heterogeneity in PET distribution
  • Measured fluorescence of TPA on plates, unable to quantify LC cutinase activity

Harvard BioDesign
  • Petase function confirmed with PNPB
  • Bacteria produced electric current when supplied with unspecified quantity of TPA

Tianjin
  • EM confirmation of PETase activity of PET film degradation
  • Multispectral scanning quantified PETase products for cell free system

TJUSLS China
  • HPLC detection of MHET to confirm PETase activity in varying conditions
  • Surface display of PETase in E. coli

UESTC-China
  • SEM and PNPB to confirm PETase activity
  • Possible detection of TPA by UV vis (higher absorbance across spectrum)

UoA New Zealand
  • Assembled PETase part with His tag

2017
Baltimore Bio-Crew
  • Fluorescine diacetate hydrolysis assay to confirm PETase and MHETase hydrolytic activity

BOKU-Vienna
    Discussion of a possible method for directed evolution of PETase by culturing cells on PET film that fluoresces when degraded

ITB Indonesia
  • Successful biofilm formation on PET, but biofilm matrix hampered PETase activity
  • Engineered E. coli ethylene glycol metabolism with directed evolution

2018
Makerere University
  • TBA

OLS Canmore Canada
  • TBA

RHIT
  • TBA

ULaVerne
  • TBA

UMaryland
  • Check out our results on this wiki!

Yale
  • TBA

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