Difference between revisions of "Team:UMaryland/PETaseIntro"
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| − | + | - Confirmed cutinase activity using PNPB esterase assay | |
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| − | + | - Engineered E. coli ethylene glycol metabolism with directed evolution | |
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| − | + | - Isolation of cutinase gene from nature with primers | |
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| − | Surface display of cutinase on E. coli | + | - Surface display of cutinase on E. coli |
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| − | Attempted TPA transport into E. coli, further research required | + | - Attempted TPA transport into E. coli, further research required |
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| − | Expressed all TPH enzymes, did not attempt to measure activity | + | - Expressed all TPH enzymes, did not attempt to measure activity |
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| − | Confirmed anaerobic conversion of PCA via AroY and XylE enzymes | + | - Confirmed anaerobic conversion of PCA via AroY and XylE enzymes |
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| − | Produced P3HB bioplastic from mixed waste containing at least some PET | + | - Produced P3HB bioplastic from mixed waste containing at least some PET |
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| − | Reduced catechol to pyruvate | + | - Reduced catechol to pyruvate |
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| − | LC cutinatse activity confimed with SEM, PNPB | + | - LC cutinatse activity confimed with SEM, PNPB |
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| − | PNPB assay to confirm activity of esterase EST13 | + | - PNPB assay to confirm activity of esterase EST13 |
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| − | Fluorescent detection of TPA can not be accomplished when in LB broth. | + | - Fluorescent detection of TPA can not be accomplished when in LB broth. |
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| − | Petase function confirmed with PNPB | + | - Petase function confirmed with PNPB |
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| − | Bacteria produced electric current when supplied with unspecified quantity of TPA | + | - Bacteria produced electric current when supplied with unspecified quantity of TPA |
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| − | Attempt at detecting PET degradation by mass change failed | + | - Attempt at detecting PET degradation by mass change failed |
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| − | Assembled PETase part with His tag | + | - Assembled PETase part with His tag |
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| − | PNPB and EM to confirm LC cutinase activity | + | - PNPB and EM to confirm LC cutinase activity |
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| − | 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. |
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| − | 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. | + | - 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. |
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| − | Unable to determine enzyme efficiency based on growth due to heterogeneity in PET distribution | + | - Unable to determine enzyme efficiency based on growth due to heterogeneity in PET distribution |
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| − | Measured fluorescence of TPA on plates, unable to quantify LC cutinase activity. | + | - Measured fluorescence of TPA on plates, unable to quantify LC cutinase activity. |
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| − | HPLC detection of MHET to confirm PETase activity in varying conditions | + | - HPLC detection of MHET to confirm PETase activity in varying conditions |
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| − | Surface display of PETase in E. coli | + | - Surface display of PETase in E. coli |
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| − | EM confirmation of PETase activity of PET film degradation | + | - EM confirmation of PETase activity of PET film degradation |
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| − | Multispectral scanning quantified PETase products for cell free system | + | - Multispectral scanning quantified PETase products for cell free system |
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| − | Planned to weigh PET degradation, no results | + | - Planned to weigh PET degradation, no results |
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| − | SEM and PNPB to confirm PETase activity | + | - SEM and PNPB to confirm PETase activity |
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| − | Possible detection of TPA by UV vis (higher absorbance across spectrum) | + | - Possible detection of TPA by UV vis (higher absorbance across spectrum) |
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| − | Withdrawn | + | - Withdrawn |
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| − | PNPB and SEM to confirm PETase activity | + | - PNPB and SEM to confirm PETase activity |
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| − | Successful biofilm formation on PET, but biofilm matrix hampered PETase activity. | + | - Successful biofilm formation on PET, but biofilm matrix hampered PETase activity. |
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| − | Fluorescine diacetate hydrolysis assay to confirm PETase and MHETase hydrolytic activity | + | - Fluorescine diacetate hydrolysis assay to confirm PETase and MHETase hydrolytic activity |
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| − | 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 |
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| + | ________________________________________________________________________________________________ | ||
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Revision as of 16:31, 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.