Difference between revisions of "Part:BBa K3722008"

 
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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.
 
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.
  
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===Usage and Biology===
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===Biology===
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MaFMO hydroxylates the C3 position of the indole ring and forms 6-Br-indoxyl.
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<html>
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    <figure>
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        <img src="https://2021.igem.org/wiki/images/9/9d/T--NWU-CHINA-A--part081.png" width="80%" style="float:center">
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        <figcaption>
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        <p style="font-size:1rem">
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        </p>
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        </figcaption>
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    </figure>
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</html>
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We used it to oxidate 6-Br-indole and produce 6,6’-dibromoindigo(Tyrian purple).
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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.
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 +
 
 +
 
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===Characterization===
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The verification of the expression
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We used IPTG to induce the expression at different times.
 +
<html>
 +
    <figure>
 +
        <img src="https://2021.igem.org/wiki/images/2/2b/T--NWU-CHINA-A--part084.png" width="80%" style="float:center">
 +
        <figcaption>
 +
        <p style="font-size:1rem">
 +
        </p>
 +
        </figcaption>
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    </figure>
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</html>
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:'''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)
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===References===
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Han, G. H., Shin, H. J. & Kim, S. W . Optimization of bio-indigo productionby recombinant E-coli harboring fmo gene. Enzyme. Micro. Tech. 42, 
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617–623 (2008).
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Revision as of 00:23, 22 October 2021


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]