Part:BBa_K3722008

T7_Promoter-RBS-MaFMO-T7_Terminator

An oxidation reaction enzyme that catalyzes the binding of oxygen atoms of molecular oxygen to the substrate，Monooxygenase hydroxylates the C3 position of the indole ring and forms 6-Br-indoxyl.

Biology

MaFMO hydroxylates the C3 position of the indole ring and forms 6-Br-indoxyl.

We used it to oxidate 6-Br-indole and produce 6,6’-dibromoindigo(Tyrian purple). In addition, MaFMO can oxidized indole to indigo.

Usage

We construct the composite part BBa_K3332052 and transformed it into E. coli BL21 (DE3) to verify its expression.

Characterization

The verification of the expression We used IPTG to induce the expression at different times.

Fig.1 SDS-PAGE analysis of MaFMO .Induction condition: LB media, OD600 0.6 ~ 0.8, IPTG 0.1 mM, 18°C and 200 rpm.(0:negitive control 1:4h 2:5h 3:6h 4:7h 5:8h 6:9h 7:10h 8:11h)

References

Han, G. H., Shin, H. J. & Kim, S. W . Optimization of bio-indigo productionby recombinant E-coli harboring fmo gene. Enzyme. Micro. Tech. 42, 617–623 (2008).

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 1435
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal XhoI site found at 633
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 213
  Illegal AgeI site found at 236
  Illegal AgeI site found at 870
  
• 1000
  COMPATIBLE WITH RFC[1000]