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

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]