Part:BBa_K2982005:Design

Coding sequence for 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, Isoleucine 253 residues are located at the corresponding positions of Serine 245 in subsite II of IsPETase. The resulting double mutant makes the substrate binding site, subunit 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