Part:BBa_K5157011

PETase

Usage and Biology

IsPETase, a PET hydrolase discovered from the bacterium Ideonella sakaiensis 201-F6, has become a research favorite for its ability to efficiently degrade PET at low temperature ^[1]. It breaks PET into bis(2-hydroxyethyl) terephthalate (BHET) and 2-hydroxyethyl methyl terephthalate (MHET) by hydrolyzing the ester bond, followed by hydrolysis to the end products EG and TPA with the help of MHETase (Fig. 1).

Fig. 1. Schematic diagram of the PETase pathway for PET degradation.

The enzyme we have chosen is a IsPETase-derived modified mutant with high catalytic performance and stability, obtained from a previous study by our research group. The enzyme sequence has been codon optimized for E. coli. We would construct a fusion protein containing this sequence and characterize it by chassis cell E. coli BL21 (DE3).

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal SpeI site found at 751
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal SpeI site found at 751
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 614
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal SpeI site found at 751
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal SpeI site found at 751
  Illegal NgoMIV site found at 142
  
• 1000
  COMPATIBLE WITH RFC[1000]

References

[1] Yoshida S, Hiraga K, Takehana T, et al. A bacterium that degrades and assimilates poly(ethylene terephthalate) [J]. Science, 2016, 351(6278): 1196-1199.