Part:BBa_K1067003

Ammonium monooxygenase from Nitrosomonas europaea

Ammonia monooxygenase (AMO) from Nitrosomonas europaea converts ammonia to hydroxylamine;

NH₃ + O₂ + cycAX_red + 2 H⁺ → NH₂OH + H₂O + cycAX_ox

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

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE 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
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 2028
1000
COMPATIBLE WITH RFC[1000]

Characterization

File: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.