Coding

Part:BBa_K2982001:Design

Designed by: Tsang Hoi Yeung, Ocean   Group: iGEM19_HK_GTC   (2019-08-03)


Coding sequence for W159H/S238F 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 double mutant narrows active site to make it more TfCutinase-like which increases its activity on PET plastic


Source

Sequence of double mutant in '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