Difference between revisions of "Part:BBa K4229016"

Revision as of 23:37, 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 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

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

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K4229016 short</partinfo>
-These are the genes we put inside MCS1 of our vector pCDF-Duet-1. The first gene is the fusion protein of TnaA with the snoopcatcher(BBa_K4229013) on the C-terminal, the second one being Fre(BBa_K4229001). It was decided to put the enzymes with the catcher at the beginning of each MCS as those are our most important proteins, which we wanted to have produced the most. These two genes build together with the Biobrick BBa_K42218 all enzymes of the indigo/indirubin pathway. The production was tested under the T7 promotor and LacI operator in those two Biobricks: BBa_K4229033 and BBa_K4229034)
+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.
 [[File:Grafik indigo weier bg.png|900px|thumb|left|]]
-<!-- Add more about the biology of this part here
+<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].
 ===Usage and Biology===
+The production was tested under the T7 promoter and LacI operator in those two Biobricks: BBa_K4229033 and BBa_K4229034)
+<b>References</b>
+<br>
+[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.
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