Team:UMaryland/PETaseIntro
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Withdrawn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.