Part:BBa_K3039019

PETase Reconstructed Ancestor 3

The enzymes PETase and MHETase were first discovered in Ideonella sakaiensis in 2016 by a group of researchers in Japan. These enzymes were found to degrade polyethylene terephthalate (PET) into its monomers, terephthalic acid (TPA) and ethylene glycol (EG). PETase degrades PET into Mono-(2-hydroxyethyl)terephthalic acid (MHET), Bis(2-Hydroxyethyl) terephthalate (BHET) and TPA, the main product being MHET. MHET is further degraded by MHETase into TPA and EG. We are aiming to use mutants of these enzymes to degrade the microfibres that are coming off clothing during washing cycles. The enzymes would be secreted into a filter that captures the microfibres. This is the sequence of one of the four reconstructed ancestors of PETase with a His tag attached to it. The sequence has been obtained through the method of ancestral reconstruction. The His tag has been used in order to more easily identify the enzyme.

Expression in E.coli

Assays

PET Assay

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 255
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 255
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 255
  Illegal BamHI site found at 21
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 255
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 255
  
• 1000
  COMPATIBLE WITH RFC[1000]