Part:BBa_K2088010

A device that expresses the XylE- catechol 2,3-dioxygenase to degrade the catechol .

In this device, promoter J23100 and the RBS B0034 are connected before the XylE - catechol 2,3-dioxygenase.

Catechol is one of the main metabolites during the degradation of aromatic compounds. This device is useful to degrade catechol in some degradation systems to reduce the toxicity of catechol to cells. With the strong promoter and rbs, xylE can convert to 2-hydroxymuconate semialdehyde, a bright yellow product which can be metabolized by E.coli.

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 400
  Illegal NgoMIV site found at 572
  Illegal AgeI site found at 923
  
• 1000
  COMPATIBLE WITH RFC[1000]

Qualitative Experiment

• We verified the function of xylE by constructing a device BBa_K2088010.

Figure 8: We construced a device that includes J23100, B0034, xylE and B0014. Transform the device into E.coli and culture at 37℃ overnight, the colonies showed a common white colour.

Figure 9: Drip 0.2mol/L catechol onto the colonies, the cells became bright yellow after 10 minutes. This indicates the xylE converts catechol to 2-hydroxy-cis,cis-muconic semialdehyde.