Difference between revisions of "Part:BBa K4390073"
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FAST-PETase is a mutant form of the PETase, which has faster activity. We immobilised this part to Silica beads as part of our cell free PET biodegradation device.
  
 
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Revision as of 02:32, 12 October 2022


FAST-PETase

FAST-PETase is a mutant form of the PETase, which has faster activity. We immobilised this part to Silica beads as part of our cell free PET biodegradation device.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 139
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 368

Improvement on BBa_K3946023 by Edinburgh-UHAS_Ghana 2022

We improved upon the Dou-PETase part (Part:BBa_K3946023) with the FAST-PETase part (Part:BBa_K4390073).

PETase-silica tag fusion protein Activity Test

We assessed the various PETase activities using a para-nitrophenol-butyrate (pNPB) assay, since PETases can hydrolyse pNPB into para-nitrophenol (pNP), which strongly absorbs at 415 nm. pNPB is not the PETase’s true substrate, but this preliminary assay is still representative of PETase activity. Figure 1 shows that untagged FAST-PETase has higher activity than Dou-PETase in this assay. Hence, FAST-PETase is an improvement over Dou-PETase for degradation of PET. The data also shows that PETase activity is diminished, but still exists when immobilised on silica beads, as the activity is higher than SHuffle E. coli lysate.

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