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1.7.3.1: nitroalkane oxidase

This is an abbreviated version!
For detailed information about nitroalkane oxidase, go to the full flat file.

Word Map on EC 1.7.3.1

Reaction

ethylnitronate
+
O2
+
FMNH2
=
acetaldehyde
+
nitrite
+
FMN
+
H2O

Synonyms

2-npdl, More, NAO, NaoA, nitroalkane oxidase, nitroalkane-oxidizing enzyme, nitroethane oxidase, nitroethane:oxygen oxidoreductase, NOE, oxidase, nitroethane, PA4202

ECTree

     1 Oxidoreductases
         1.7 Acting on other nitrogenous compounds as donors
             1.7.3 With oxygen as acceptor
                1.7.3.1 nitroalkane oxidase

Crystallization

Crystallization on EC 1.7.3.1 - nitroalkane oxidase

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CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
crystallization data of mutant D402N in complex with 1-nitrohexane or 1-nitrooctane show the presence of substrate in the binding site. The aliphatic chain of the substrate extends into a tunnel leading to the enzyme surface. The oxygens of the substrate nitro group interact both with amino acid residues and with the 2'-hydroxyl of the FAD. The structure of wild-type enzyme trapped with cyanide during oxidation of 1-nitrohexane shows the presence of the modified flavin. A continuous hydrogen bond network connects the nitrogen of the CN-hexyl-FAD through the FAD 2'-hydroxyl to a chain of water molecules extending to the protein surface. Data for mutant S276A in complex with nitrohexane
hanging drop vapour diffusion method
-
hexagonal rod-shaped crystals of R409K and D402E NAO were obtained using hanging drop vapor-diffusion methods
-
purified recombinant wild-type enzyme, crystallization of the native enzyme in 2 different crystal forms and of the selenomethionine-labeled enzyme in a third one, hanging drop vapour diffusion method, sodium cacodylate buffer, pH 7.5, containing spermidine hydrochloride, and PEG 4000 at varying concentrations for all 3 mixtures, crystal form 2 requires addition of 1,6-hexanediol at 8% w/v, crystal form 3 requires DTT at 10 mM, 4°C, 10-14 days, X-ray diffraction structure determinations and analysis at 3.2-2.0 A resolution or below, three-wavelength MAD data
-
vapor diffusion method, using 2.5 M magnesium sulfate, 0.1 M 2-(N-morpholino) ethanesulfonic acid buffer, pH 6.0, and 18% (v/v) glycerol
mutant enzyme H183S in complex with nitroethane and FMN, hanging drop vapor diffusion method, using 0.2 M ammonium fluoride, 20% (w/v) polyethylene glycol 3350
-
native and a selenomethionine-substituted enzyme, to 1.9 A resolution. Primitive orthorhombic space group P21, with unit-cell parameters a = 70.06, b = 55.43, c = 87.74 A, beta = 96.56° for native NAO and a = 69.89, b = 54.83, c = 88.20 A, beta = 95.79° for selenomethionine-substituted enzyme
-
wild-type and mutant H179D. Enzyme consists of two domains, a TIM barrel domain bound to FMN and C-terminal domain with a alpha-alpha-alpha-beta-alpha-beta-alpha fold. It shows the typical function as nitroalkane oxidase but its structure is similar to that of 2-nitropropane dioxygenase