Difference between revisions of "Part:BBa K2982004:Design"

 
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===Source===
 
===Source===
  
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Modified from wild type IsPETase sequence from 'Structural Insight into Molecular Mechanism of Poly (ethylene terephthalate) Degradation', Nature Communication, 2018.
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Modified from wild type IsPETase sequence from 'Characterization and engineering of a plastic-degrading aromatic polyesterase', PNAs, 2018.
  
 
===References===
 
===References===
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Austin, H. P., Allen, M. D., Donohoe, B. S., Rorrer, N. A., Kearns, F. L., Silveira, R. L., . . . Beckham, G. T. (2018). Characterization and engineering of a plastic-degrading aromatic polyesterase. Proceedings of the National Academy of Sciences, 115(19). doi:10.1073/pnas.1718804115

Latest revision as of 04:40, 7 August 2019


Coding sequence for W159H/S245I IsPETase double mutant


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 348
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 304
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 348
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 348
    Illegal AgeI site found at 627
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The mutation sites locate in substrate binding site, subsite II where three MHET moieties are bound through hydrophobic interaction. In TfCut2, Histidine 169 residues and Isoleucine 253 are located at the corresponding positions of Trpytophan 159 and Serine 245 in subsite II of IsPETase. The resulting double mutant makes the substrate binding site, substrate II more cutinase-like and increases the hydrophobic property of the enzyme.


Source

Modified from wild type IsPETase sequence from 'Characterization and engineering of a plastic-degrading aromatic polyesterase', PNAs, 2018.

References

Austin, H. P., Allen, M. D., Donohoe, B. S., Rorrer, N. A., Kearns, F. L., Silveira, R. L., . . . Beckham, G. T. (2018). Characterization and engineering of a plastic-degrading aromatic polyesterase. Proceedings of the National Academy of Sciences, 115(19). doi:10.1073/pnas.1718804115