Part:BBa_K4390073

FAST-PETase

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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.