Coding

Part:BBa_K5023003

Designed by: Lucas Daniel Ovelar Vargas   Group: iGEM23_UNILA-LatAm   (2023-10-08)
Revision as of 20:21, 8 October 2023 by Lucasovelar (Talk | contribs)

FAST-PETase

FAST-PETase is a mutant variant derived from the PETase enzyme. It contains five mutations compared to the wild-type PETase. These mutations include N233K, R224Q, and S121E from prediction, along with D186H and R280A from the scaffold. This enzyme demonstrated superior PET-hydrolytic activity relative to both wild-type and other engineered alternatives between 30 and 50°C across a range of pH levels. At 50°C, FAST-PETase showed the highest overall degradation of all mutants tested, releasing 33.8 mM of PET monomers (the sum of terephthalic acid (TPA) and mono-(2-hydroxyethyl) terephthalate (MHET)). The enzyme's activity against post-consumer PET (pc-PET) was substantially higher than that of other enzymes like WT PETase, ThermoPETase, DuraPETase, LCC, and ICCM under the same conditions. FAST-PETase was able to almost completely degrade untreated post-consumer PET from 51 different thermoformed products in just one week. The enzyme also demonstrated the capability to depolymerize untreated, amorphous portions of a commercial water bottle. A time-course analysis revealed an almost linear decay rate of PET degradation and a concomitant increase in crystallinity over time. The development of FAST-PETase offers a potential solution for the degradation of PET plastics, especially given its enhanced activity and stability across a range of conditions. This enzyme can play a pivotal role in addressing the environmental challenges posed by PET plastic accumulation.   Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 846
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 846
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 351
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 846
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 846
    Illegal NgoMIV site found at 155
  • 1000
    COMPATIBLE WITH RFC[1000]


Reference=

Lu, H., Diaz, D.J., Czarnecki, N.J. et al. Machine learning-aided engineering of hydrolases for PET depolymerization. Nature 604, 662–667 (2022). https://doi.org/10.1038/s41586-022-04599-z

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