Difference between revisions of "Part:BBa K1067003"
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| − | Ammonia consumption with respect to growth (optical density) for two replicates of the AMO transformant, and untransformed E. coli control. The rate of consumption of ammonia is much higher in the transformant than in the control. [https://static.igem.org/mediawiki/2013/e/ed/Dtu_Amo_confirmation.png | + | Ammonia consumption with respect to growth (optical density) for two replicates of the AMO transformant, and untransformed E. coli control. The rate of consumption of ammonia is much higher in the transformant than in the control. [https://static.igem.org/mediawiki/2013/e/ed/Dtu_Amo_confirmation.png (larger image)] |
Revision as of 10:14, 12 October 2013
Ammonium monooxygenase from Nitrosomonas europaea
Ammonia monooxygenase (AMO) from Nitrosomonas europaea converts ammonia to hydroxylamine;
NH3 + O2 + cycAXred + 2 H+ → NH2OH + H2O + cycAXox
The whole operon containing amoA1, amoB1, amoC1 http://ecocyc.org/NEUR228410/NEW-IMAGE?type=MAPTICK&object=2238597/2241670&chromosome=CHROMOSOME-1&bp-range=2238597/2241670 link to Biocyc was extracted by colony PCR on a N. europaea culture.
Usage and Biology
AMO is an integral membrane protein composed of 3 subunits (amoA, amoB, amoC) and is first step in the enzymatic conversion of ammonia to nitrite in the nitrifying bacteria genus. The protein works in close relationship with cycA, cycX and Hydroxylamine oxidoreductase (HAO) in this process. HAO is providing 2 free electrons that are picked up and delivered to AMO by the cytochromes cycA and cycX. The electron from HAO is what makes AMO's work possible.
This part is experienced to slow down growth rates of E.coli substantially; likely because this is a heterologous expressed membrane protein and thereby disrupts E.coli membrane.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 263
Illegal BamHI site found at 306
Illegal BamHI site found at 2510
Illegal BamHI site found at 2822 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 2028
- 1000COMPATIBLE WITH RFC[1000]
Characterization
Ammonia consumption with respect to growth (optical density) for two replicates of the AMO transformant, and untransformed E. coli control. The rate of consumption of ammonia is much higher in the transformant than in the control. (larger image)
