Part:BBa_K4229016
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 Biobrick 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
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 990
- 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 990
- 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 990
- 25INCOMPATIBLE 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 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 2150
Illegal SapI.rc site found at 2270
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