Difference between revisions of "Team:UMaryland/PETaseIntro"

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SUMMARY OF WET LAB RESULTS
 
SUMMARY OF WET LAB RESULTS
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UC DAVIS 2012
 
UC DAVIS 2012
Confirmed cutinase activity using PNPB esterase assay
+
<p>
Engineered E. coli ethylene glycol metabolism with directed evolution
+
<p>
 +
        Confirmed cutinase activity using PNPB esterase assay
 +
<p>
 +
<p>
 +
        Engineered E. coli ethylene glycol metabolism with directed evolution
 +
<p>
 +
<p>
 
BAU-Indonesia 2012
 
BAU-Indonesia 2012
Isolation of cutinase gene from nature with primers
+
<p>
 +
<p>
 +
        Isolation of cutinase gene from nature with primers
 +
<p>
 +
<p>
 
TU Darmstadt 2012
 
TU Darmstadt 2012
 +
<p>
 +
<p>
 
Surface display of cutinase on E. coli
 
Surface display of cutinase on E. coli
 +
<p>
 +
<p>
 
Attempted TPA transport into E. coli, further research required
 
Attempted TPA transport into E. coli, further research required
 +
<p>
 +
<p>
 
Expressed all TPH enzymes, did not attempt to measure activity
 
Expressed all TPH enzymes, did not attempt to measure activity
 +
<p>
 +
<p>
 
Confirmed anaerobic conversion of PCA via AroY and  XylE enzymes
 
Confirmed anaerobic conversion of PCA via AroY and  XylE enzymes
 +
<p>
 +
<p>
 
Imperial_College 2013
 
Imperial_College 2013
 +
<p>
 +
<p>
 
Produced P3HB bioplastic from mixed waste containing at least some PET
 
Produced P3HB bioplastic from mixed waste containing at least some PET
 +
<p>
 +
<p>
 
METU_Turkey 2014
 
METU_Turkey 2014
 +
<p>
 +
<p>
 
Reduced catechol to pyruvate
 
Reduced catechol to pyruvate
 +
<p>
 +
<p>
 
ITB-Indonesia 2014
 
ITB-Indonesia 2014
 +
<p>
 +
<p>
 
LC cutinatse activity confimed with SEM, PNPB
 
LC cutinatse activity confimed with SEM, PNPB
 +
<p>
 +
<p>
 
Pasteur Paris 2015
 
Pasteur Paris 2015
 +
<p>
 +
<p>
 
PNPB assay to confirm activity of esterase EST13
 
PNPB assay to confirm activity of esterase EST13
 +
<p>
 +
<p>
 
Fluorescent detection of TPA can not be accomplished when in LB broth.
 
Fluorescent detection of TPA can not be accomplished when in LB broth.
 +
<p>
 +
<p>
 
Harvard 2016
 
Harvard 2016
 +
<p>
 +
<p>
 
Petase function confirmed with PNPB
 
Petase function confirmed with PNPB
 +
<p>
 +
<p>
 
Bacteria produced electric current when supplied with unspecified quantity of TPA
 
Bacteria produced electric current when supplied with unspecified quantity of TPA
 +
<p>
 +
<p>
 
ASIJ Tokyo 2016
 
ASIJ Tokyo 2016
 +
<p>
 +
<p>
 
Attempt at detecting PET degradation by mass change failed
 
Attempt at detecting PET degradation by mass change failed
 +
<p>
 +
<p>
 
UoA_NewZealand 2016
 
UoA_NewZealand 2016
 +
<p>
 +
<p>
 
Assembled PETase part with His tag
 
Assembled PETase part with His tag
 +
<p>
 +
<p>
 
BGU-Israel 2016
 
BGU-Israel 2016
 +
<p>
 +
<p>
 
PNPB and EM to confirm LC cutinase activity
 
PNPB and EM to confirm LC cutinase activity
 +
<p>
 +
<p>
 
P. putida can grow on PCA as sole carbon source, but not TPA.
 
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
+
<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.
 
Measured fluorescence of TPA on plates, unable to quantify LC cutinase activity.
 +
<p>
 +
<p>
 
TJUSLS China 2016
 
TJUSLS China 2016
 +
<p>
 +
<p>
 
HPLC detection of MHET to confirm PETase activity in varying conditions
 
HPLC detection of MHET to confirm PETase activity in varying conditions
 +
<p>
 +
<p>
 
Surface display of PETase in E. coli
 
Surface display of PETase in E. coli
 +
<p>
 +
<p>
 
Tianjin 2016
 
Tianjin 2016
 +
<p>
 +
<p>
 
EM confirmation of PETase activity of PET film degradation
 
EM confirmation of PETase activity of PET film degradation
 +
<p>
 +
<p>
 
Multispectral scanning quantified PETase products for cell free system
 
Multispectral scanning quantified PETase products for cell free system
 +
<p>
 +
<p>
 
Baltimore BioCrew 2016
 
Baltimore BioCrew 2016
 +
<p>
 +
<p>
 
Planned to weigh PET degradation, no results
 
Planned to weigh PET degradation, no results
 +
<p>
 +
<p>
 
UESTC China 2016
 
UESTC China 2016
 +
<p>
 +
<p>
 
SEM and PNPB to confirm PETase activity
 
SEM and PNPB to confirm PETase activity
 +
<p>
 +
<p>
 
Possible detection of TPA by UV vis (higher absorbance across spectrum)
 
Possible detection of TPA by UV vis (higher absorbance across spectrum)
 +
<p>
 +
<p>
 
AUC_Turkey 2016
 
AUC_Turkey 2016
 +
<p>
 +
<p>
 
Withdrawn
 
Withdrawn
 +
<p>
 +
<p>
 
ITB-Indonesia 2017
 
ITB-Indonesia 2017
 +
<p>
 +
<p>
 
PNPB and SEM to confirm PETase activity
 
PNPB and SEM to confirm PETase activity
 +
<p>
 +
<p>
 
Successful biofilm formation on PET, but biofilm matrix hampered PETase activity.
 
Successful biofilm formation on PET, but biofilm matrix hampered PETase activity.
 +
<p>
 +
<p>
 
Baltimore BioCrew 2017
 
Baltimore BioCrew 2017
 +
<p>
 +
<p>
 
Fluorescine diacetate hydrolysis assay to confirm PETase and MHETase hydrolytic activity
 
Fluorescine diacetate hydrolysis assay to confirm PETase and MHETase hydrolytic activity
 +
<p>
 +
<p>
 
BOKU-Vienna 2017
 
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
 
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.
 
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>
 
<p>

Revision as of 16:29, 15 October 2018

History

iGEM teams pursuing PET related projects

SUMMARY OF WET LAB RESULTS

UC DAVIS 2012

Confirmed cutinase activity using PNPB esterase assay

Engineered E. coli ethylene glycol metabolism with directed evolution

BAU-Indonesia 2012

Isolation of cutinase gene from nature with primers

TU Darmstadt 2012

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

Imperial_College 2013

Produced P3HB bioplastic from mixed waste containing at least some PET

METU_Turkey 2014

Reduced catechol to pyruvate

ITB-Indonesia 2014

LC cutinatse activity confimed with SEM, PNPB

Pasteur Paris 2015

PNPB assay to confirm activity of esterase EST13

Fluorescent detection of TPA can not be accomplished when in LB broth.

Harvard 2016

Petase function confirmed with PNPB

Bacteria produced electric current when supplied with unspecified quantity of TPA

ASIJ Tokyo 2016

Attempt at detecting PET degradation by mass change failed

UoA_NewZealand 2016

Assembled PETase part with His tag

BGU-Israel 2016

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.

TJUSLS China 2016

HPLC detection of MHET to confirm PETase activity in varying conditions

Surface display of PETase in E. coli

Tianjin 2016

EM confirmation of PETase activity of PET film degradation

Multispectral scanning quantified PETase products for cell free system

Baltimore BioCrew 2016

Planned to weigh PET degradation, no results

UESTC China 2016

SEM and PNPB to confirm PETase activity

Possible detection of TPA by UV vis (higher absorbance across spectrum)

AUC_Turkey 2016

Withdrawn

ITB-Indonesia 2017

PNPB and SEM to confirm PETase activity

Successful biofilm formation on PET, but biofilm matrix hampered PETase activity.

Baltimore BioCrew 2017

Fluorescine diacetate hydrolysis assay to confirm PETase and MHETase hydrolytic activity

BOKU-Vienna 2017

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

If you see this then 2018 results from other teams have not been posted in time to show them here before the wiki freeze.