Part:BBa_K3039010

SP_lamB-PETase T88A_S93M_S121E_W159F_D186H_R280A

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 sequence is the Escherichia coli K12 (E. coli K12) codon optimized DNA of the T88A_S93M_S121E_W159F_D186H_R280A mutant of PETase with a lamB signal peptide and His tag attached to it. The lamB signal peptide has been used in order to secrete the enzyme outside E.coli and the His tag is attached to more easily identify the enzymes. These mutations have been reported in past papers to increase the thermostability of PETase.

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 AgeI site found at 654
  
• 1000
  COMPATIBLE WITH RFC[1000]