Coding

Part:BBa_K2010999

Designed by: William Cho   Group: iGEM16_Harvard_BioDesign   (2016-06-17)
Revision as of 00:42, 22 October 2019 by VSajtovich (Talk | contribs)


PETase (PET-degrading enzyme, origin I. sakaiensis)

PETase is a poly(ethylene terehphthalate)-degrading enzyme first identified in Ideonella sakaiensis. BBa_K2010999 is the sequence for the E. coli K12 optimized DNA sequence for PETase. The catalytic activity of IsPETase can be characterized using a p-nitrophenol butyrate (pNPB) assay, in which PETase cleaves the ester bond of pNPB to release p-nitrophenolate. The absorbance of this compound can be measured at 405 nm to reflect IsPETase activity.

T--Toronto--WT.SDSPAGE.png

An SDS-PAGE gel demonstrates successful protein purification of PETase, which has a molecular weight of approximately 30 kDa.

T--Toronto--pNPB-Hydrolysis-pH-7_Wildtype-All-Conc.png

This graph demonstrates PETase activity with various substrate concentrations at a pH of 7.0.

T--Toronto--pNPB-Hydrolysis-pH-8-Wildtype-All-Conc.png

This graph demonstrates PETase activity with various substrate concentrations at a pH of 8.0.

T--Toronto--pNPB-Hydrolysis-pH-9_Wildtype-All-Conc.png

This graph demonstrates PETase activity with various substrate concentrations at a pH of 9.0.

T--Toronto--pNPB-Hydrolysis-Wildtype_pH7-8-9.png

This graph demonstrates PETase activity as a function of pH. PETase is most active at a pH of 9.0.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NotI site found at 67
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
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