Difference between revisions of "Part:BBa K4229016"
 
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<b>Figure 1: Schematic representation of the indigo/indirubin pathway.</b> L-tryptophan is imported by the membrane protein TnaB (low affinity tryptophan permease). L-tryptophan is cleaved into indole, NH4+ and pyruvate by the tryptophanase TnaA. The reaction continues by the hydroxylation of indole through XiaI. To enhance the effectivity of this enzyme, the NAD(P)H-flavin reductase provides XiaI with FADH2 by adding hydrogen to FAD. Finally, indole is transformed to either 3-Hydroxyindole or 2-Hydroxyindole. These two substances spontaneously react to 3-Oxindole and 2-Oxindole through the secession of hydrogen from the OH-group. Through spontaneous dimerization indigo and indirubin are formed. Graphic adapted from [1].]]  
 
<b>Figure 1: Schematic representation of the indigo/indirubin pathway.</b> L-tryptophan is imported by the membrane protein TnaB (low affinity tryptophan permease). L-tryptophan is cleaved into indole, NH4+ and pyruvate by the tryptophanase TnaA. The reaction continues by the hydroxylation of indole through XiaI. To enhance the effectivity of this enzyme, the NAD(P)H-flavin reductase provides XiaI with FADH2 by adding hydrogen to FAD. Finally, indole is transformed to either 3-Hydroxyindole or 2-Hydroxyindole. These two substances spontaneously react to 3-Oxindole and 2-Oxindole through the secession of hydrogen from the OH-group. Through spontaneous dimerization indigo and indirubin are formed. Graphic adapted from [1].]]  
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Latest revision as of 23:51, 11 October 2022


RBS-snoopCatcherTnaA-RBS-FRE

This composite part was used to for the coexpression of two target genes of the indigo/indirubin pathways. The first one is TnaA with N-terminal SnoopCatcher (BBa_K4229013) and the second one being Fre (BBa_K4229001). These two genes build together with the BBa_K4229018 all enzymes of the indigo/indirubin pathway.

Figure 1: Schematic representation of the indigo/indirubin pathway. L-tryptophan is imported by the membrane protein TnaB (low affinity tryptophan permease). L-tryptophan is cleaved into indole, NH4+ and pyruvate by the tryptophanase TnaA. The reaction continues by the hydroxylation of indole through XiaI. To enhance the effectivity of this enzyme, the NAD(P)H-flavin reductase provides XiaI with FADH2 by adding hydrogen to FAD. Finally, indole is transformed to either 3-Hydroxyindole or 2-Hydroxyindole. These two substances spontaneously react to 3-Oxindole and 2-Oxindole through the secession of hydrogen from the OH-group. Through spontaneous dimerization indigo and indirubin are formed. Graphic adapted from [1].




Usage and Biology

The production was tested under the T7 promoter and LacI operator in those two Biobricks: BBa_K4229033 and BBa_K4229034)


References
[1] H. Yin et al., “Efficient Bioproduction of Indigo and Indirubin by Optimizing a Novel Terpenoid Cyclase XiaI in Escherichia coli,” ACS Omega, vol. 6, no. 31, pp. 20569–20576, 2021, doi: 10.1021/acsomega.1c02679.


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 990
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 990
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 990
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 990
    Illegal AgeI site found at 155
    Illegal AgeI site found at 241
    Illegal AgeI site found at 1613
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 2150
    Illegal SapI.rc site found at 2270