Part:BBa_K2228000
Benzene Degradation Upper Operon
The part contains a TetR coding region under the control of a constitutive promoter and an operon under the control of a TetR promoter. The operon encodes for a three enzyme- six gene pathway that converts benzene to 2-hydroxymuconate-6-semialdehyde, going through cis-1,2-dihydrobenzene-1,2-diol and catechol as intermediates. This pathway can then be combined with a separate pathway (device unsubmitted) to convert 2-hydroxymuconate-6-semialdehyde to the final products of pyruvate and acetyl-CoA. The enzymes used are benzene-1,2 dioxygenase, cis-1,2-dihydrobenzene-1,2-diol dehydrogenase, and catechol-2,3-dioxygenase. We refer to this pathway as the "upper operon," and the unsubmitted pathway as the "lower operon," which together degrade benzene for incorporation into metabolism. The whole pathway was split into two operons due to their large size. For details about sequencing, protein expression, and characterization, please view http://2017.igem.org/Team:Purdue/Experiments/Benzene_Operon
The benzene degradation pathway has been found in the Rhodococcus strain 91, Rhodococcus strain332, and Escherichia fergusonii.3a
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
aThis was contributed by the 2020 University of Newcastle Australia iGEM Team, the DeNovocastrians
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30
Illegal NheI site found at 1899 - 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 942
Illegal BamHI site found at 5635
Illegal XhoI site found at 1210 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE 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 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1543
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