Coding

Part:BBa_K2910003

Designed by: Dimitrije Ratkov   Group: iGEM19_Toronto   (2019-10-16)
Revision as of 03:27, 22 October 2019 by AdrianaDLP (Talk | contribs)


IsPETase-T88F-I208Y-S238F-S245Y-R280A with C-terminal Hexahistidine Tag

This part encodes Ideonella sakaiensis poly(ethylene terephthalate) hydrolase with several mutations predicted to increase its catalytic activity. The bulky arginine residue at 280 has been mutated to an alanine to reduce steric clash with PET. The other mutations are predicted to increase interaction between isPETase and aromatic moieties of PET.

Internal working name: Dimi1

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]

A plot of pNB hydrolysis by BBa_K2910003 at pH = 8, concentration = 2500uM; is shown below:

<img scr="T--Toronto--pNPB-Hydrolysis-pH-8_Dimi-2500uM.png">


A plot of pNB hydrolysis by BBa_K2910003 at pH = 8, concentration = 625 uM; is shown below:

<img scr="T--Toronto--pNPB-Hydrolysis-pH-8_Dimi-625uM.png">


A plot of pNB hydrolysis by BBa_K2910003 at pH = 8, concentration = 156uM; is shown below:

<img scr="T--Toronto--pNPB-Hydrolysis-pH-8_Dimi-156uM.png">


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Parameters
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