Coding

Part:BBa_K2910006

Designed by: Dimitrije Ratkov   Group: iGEM19_Toronto   (2019-10-16)


IsPETase-T88I-I208Y-S238F-N241T-S245Y-R280T 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 threonine, to reduce steric clash with PET and to contribute a hydrogen-bonding interaction with ester oxygen in the PET chain. Mutations at sites 208, 238, and 245 are predicted to increase interaction between isPETase and aromatic moieties of PET. T88 was mutated to isoleucine to better interact with a nonpolar PET moiety. N241 was mutated to threonine, to better-position a hydrogen-bonding interaction with ester oxygen in the PET chain.

Internal working name: Dimi4

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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