Part:BBa_K3887001:Design

SttH

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 178
  Illegal EcoRI site found at 550
  Illegal PstI site found at 337
  Illegal PstI site found at 436
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 178
  Illegal EcoRI site found at 550
  Illegal PstI site found at 337
  Illegal PstI site found at 436
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 178
  Illegal EcoRI site found at 550
  Illegal XhoI site found at 1081
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 178
  Illegal EcoRI site found at 550
  Illegal PstI site found at 337
  Illegal PstI site found at 436
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 178
  Illegal EcoRI site found at 550
  Illegal PstI site found at 337
  Illegal PstI site found at 436
  Illegal NgoMIV site found at 210
  Illegal NgoMIV site found at 468
  Illegal NgoMIV site found at 1171
  Illegal NgoMIV site found at 1437
  Illegal AgeI site found at 356
  Illegal AgeI site found at 697
  
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

SttH is an enzyme from Streptomyces toxytricini used during the conversion of indole. In our experiment, SttH and TnaA to convert Trp to 6-Br-Trp, then to 6-Br-indole. The strength of SttH is it shows strict regiospecificity for both bromination and chlorination. This made it suitable for the bromination of indole. However, SttH alone shows low reactivity and exists insoluble form, so it was expressed with Fre to increase the conversion of Trp to 6-Br-Trp.

Experiment shows by using 2.5mM of indole or Trp, 0.29mM of 6-Br-indole or 0.78mM of 6-Br-Trp is produced, respectively. SttH is 2.5 times more active toward Trp than to indole. So, the SttH reaction from Trp to 6-Br-Trp is followed by TnaA converting 6-Br-Trp to 6-Br-indole.

Source

Trp 6-halogenase from Streptomyces toxytricini (SttH)