Part:BBa_K4229015
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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 879
Illegal AgeI site found at 769 - 1000COMPATIBLE WITH RFC[1000]
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