Device

Part:BBa_K2228000:Design

Designed by: Caleigh Roleck   Group: iGEM17_Purdue   (2017-10-27)


Benzene Degradation Upper Operon


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
    Illegal NheI site found at 1899
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 942
    Illegal BamHI site found at 5635
    Illegal XhoI site found at 1210
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1013
    Illegal NgoMIV site found at 3067
    Illegal NgoMIV site found at 4749
    Illegal NgoMIV site found at 6266
    Illegal AgeI site found at 50
    Illegal AgeI site found at 1045
    Illegal AgeI site found at 3099
    Illegal AgeI site found at 4781
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1543


Design Notes

The goal of our project was to degrade inhaled carcinogens, so the initial substrate (benzene) must have strong evidence for being a carcinogen and must be commonly inhaled. Further, the final end products of the whole pathway must be incorporated into metabolism. While 2-hydroxymuconate-6-semialdehyde does not incorporate into metabolism, a second pathway can then convert 2-hydroxymuconate-6-semialdehyde to pyruvate and acetyl-CoA

The benzene degradation pathway has been found in the Rhodococcus strain 91, Rhodococcus strain332, and Escherichia fergusonii.3a



aThis was contributed by the 2020 University of Newcastle Australia iGEM Team, the DeNovocastrians


Source

The TetR protein comes from the registry (BBa_C0040). The 4 genes for the benzene-1,2 dioxygenase BnzA, BnzB, BnzC, and BnzD) and the cis-1,2-dihydrobenzene-1,2-diol dehydrogenase gene (BnzE) come from Pseudomonas putida. The catechol-2,3-dioxygenase gene (CatE) comes from Bacillus subtilis.

References

1. Subashchandrabose, S., Venkateswarlu, K., Naidu, R., & Megharaj, M. (2019). Biodegradation of high-molecular weight PAHs by Rhodococcus wratislaviensis strain 9: Overexpression of amidohydrolase induced by pyrene and BaP. Science Of The Total Environment, 651, 813-821. doi: 10.1016/j.scitotenv.2018.09.192 2. Gutiérrez, T., Learmonth, R., & Couperwhite, I. (2009). Analysis of Benzene-Induced Effects onRhodococcussp. 33 Reveals that Constitutive Processes Play a Major Role in Conferring Tolerance. The Scientific World JOURNAL, 9, 209-223. doi: 10.1100/tsw.2009.29 3. Mukherjee, S., De, A., Sarkar, N., & Saha, N. (2019). Aerobic Degradation of Benzene by Escherichia spp. from Petroleum-contaminated Sites in Kolkata, West Bengal, India. Journal Of Pure And Applied Microbiology, 13(4), 2353-2362. doi: 10.22207/jpam.13.4.51