Difference between revisions of "Part:BBa K3887001:Design"

 
 
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===Design Notes===
 
===Design Notes===
−
Bromo for indole and Tryptophan
+
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===
 
===Source===

Latest revision as of 21:57, 16 October 2021


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)


References