Difference between revisions of "Part:BBa K4229015"

Revision as of 23:21, 11 October 2022

spyCatcherXiaI

XiaI is a novel terpenoid cyclase, which was taken out of the xiamycin gene cluster in Streptomyces sp. XiaI turnes indol into either 3-Oxindole or 2-Oxindole. XiaI works better with the help of Fre (BBa_K4229001) to recycle NADHP and provide hydrogen to the cyclase.

This biobrick consists of XiaI, which is fused with the SpyCatcher (BBa_K4229010) on the N-terminal of the protein. With that, we made it able to be recruited into microcompartments (wiffelballs).

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].

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
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 879
Illegal AgeI site found at 769
1000
COMPATIBLE WITH RFC[1000]

@@ Line 4: / Line 4: @@
-XiaI is a novel terpenoid cyclase, which was taken out of the xiamycin gene cluster in Streptomyces sp. XiaI turnes indol into either 3-Oxindole or 2-Oxindole. XiaI workes better with the help of Fre(BBa_K4229001) wich helps to recycle NADHP and provides hydrogen to the cyclase. This biobrick consists of XiaI,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.
+XiaI is a novel terpenoid cyclase, which was taken out of the xiamycin gene cluster in <i>Streptomyces sp.</i> XiaI turnes indol into either 3-Oxindole or 2-Oxindole. XiaI works better with the help of Fre (BBa_K4229001) to recycle NADHP and provide hydrogen to the cyclase.
-[[File:Grafik indigo weier bg.png|900px|thumb|left|]]
+This biobrick consists of XiaI, which is fused with the SpyCatcher (BBa_K4229010) on the N-terminal of the protein. With that, we made it able to be recruited into microcompartments (wiffelballs).
+[[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 [1].
+<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.