Part:BBa_K1149005

pelB-pudA expression

This part can express a plastic degradation enzyme from Comamonas acidovorans that breaks down ester bonds in polyurethane. It containes a Gly-X-Ser-X-Gly motif characteristic of serine hydrolases. The sequence was codon optimized for E.coli. We added a pelB tag to ensure secretion to the N terminal and we have added a 6xpolyhistidine tag to the C terminal. The promoter is arabinose inducible.

Theoretical pathway of PUR degradation by esterase:

references:

[http://www.sciencedirect.com/science/article/pii/S0922338X99890011 Cloning and sequence analysis of a polyurethane esterase of Comamonas acidovorans 1998]

[http://www.sciencedirect.com/science/article/pii/S0964830598000663 Purification and characterization of a soluble polyurethane degrading enzyme from Comamonas acidovorans 1999]

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 125
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 65
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 529
  Illegal AgeI site found at 604
  Illegal AgeI site found at 1753
  
• 1000
  COMPATIBLE WITH RFC[1000]