Difference between revisions of "Part:BBa K4229013"

Latest revision as of 13:10, 12 October 2022

TnaA with N-terminal SnoopCatcher

TnaA belongs to the family of the tryptophanase (tna) operon leader peptide. Tryptophanase catalyses the degradation of L-tryptophan to indole, pyruvate and ammonium, enabling the bacteria to utilise tryptophan as a source of carbon, nitrogen and energy. The tna operon of E. coli contains two major structural genes, tnaA and tnaB.

SnoopCatcher (BBa_K4229009) is fused via a short N-terminal linker TnaA. The tag enables recruitment into the compartment system (wiffelball).

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 dimerisation indigo and indirubin are formed. Graphic adapted from [2].

References
[1]. A transcriptional pause synchronizes translation with transcription in the tryptophanase operon leader region. Gong F, Yanofsky C. J. Bacteriol. 185, 6472-6, (2003). View articlePMID: 14563884
[2] 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 977
12
INCOMPATIBLE WITH RFC[12]
Illegal PstI site found at 977
21
COMPATIBLE WITH RFC[21]
23
INCOMPATIBLE WITH RFC[23]
Illegal PstI site found at 977
25
INCOMPATIBLE WITH RFC[25]
Illegal PstI site found at 977
Illegal AgeI site found at 142
Illegal AgeI site found at 228
Illegal AgeI site found at 1600
1000
COMPATIBLE WITH RFC[1000]

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K4229013 short</partinfo>
-TnaA belongs to the family of tryptophanase (tna) operon leader peptide. Tryptophanase catalyses the degradation of L-tryptophan to indole, pyruvate and ammonia, enabling the bacteria to utilise tryptophan as a source of carbon, nitrogen and energy. The tna operon of E. coli contains two major structural genes, tnaA and tnaB.
+TnaA belongs to the family of the tryptophanase (tna) operon leader peptide. Tryptophanase catalyses the degradation of L-tryptophan to indole, pyruvate and ammonium, enabling the bacteria to utilise tryptophan as a source of carbon, nitrogen and energy. The tna operon of E. coli contains two major structural genes, tnaA and tnaB.
-TnaA codes for a tryptophanase. This tryptophanase turns L-tryptophan into either Indol. This biobrick consists of a version of TnaA, which is fused with the snoopcatcher(BBa_K4229009) on the N-terminal of the protein. With that, we made it able to be recruted into the wiffelball we used in our project.
+SnoopCatcher (BBa_K4229009) is fused via a short N-terminal linker TnaA. The tag enables recruitment into the compartment system (wiffelball).
-[[File:Grafik indigo weier bg.png|900px|thumb|left|]]
+[[File:Grafik indigo weier bg.png|900px|thumb|left|
-<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 [2].
+<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 dimerisation indigo and indirubin are formed. Graphic adapted from [2].]]