1.7.2.4: nitrous-oxide reductase

This is an abbreviated version!
For detailed information about nitrous-oxide reductase, go to the full flat file.

Word Map on EC 1.7.2.4

Reaction

nitrogen
+
H2O
+ 2 ferricytochrome c =
Nitrous oxide
+ 2 ferrocytochrome c + 2 H+

Synonyms

HdN2OR, McoP, MhN2OR, multicopper oxidase, N(2)OR, N2O reductase, N2OR, nitrous oxide reductase, NosZ, PpN2OR, PsN2OR, Sden_2219, SdN2OR, Z-type N2OR

ECTree

     1 Oxidoreductases
         1.7 Acting on other nitrogenous compounds as donors
             1.7.2 With a cytochrome as acceptor
                1.7.2.4 nitrous-oxide reductase

Purification

Purification on EC 1.7.2.4 - nitrous-oxide reductase

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PURIFICATION/commentary
ORGANISM
UNIPROT
LITERATURE
a pink form is isolated when the purification is done aerobically, a purple form when the purification is under anaerobic conditions
-
ammonium sulfate, DEAE-Sephadex A-50, Sephacryl S-200
-
anaerobical purification of N2OR under an Ar atmosphere by three different steps of anion exchange chromatography, followed by cytochrome c affinity chromatography and ultrafiltration
-
DEAE cellulose column chromatography, hydroxyapatite column chromatography, Mono Q10 anion-exchange column chromatography, and Sephadex S-200 gel filtration
-
DEAE-FF ion exchange chromatography and Resource Q column chromatography
-
HiTrap Q Sepharose anion exchange chromatography
-
isolation of the enzyme in the purple form, in which the copper centre is in the oxidized [2Cu2+:2Cu+] redox state and is redox active. Isolation of this form in the presence of oxygen from a microaerobic culture in the presence of nitrate and also from a strictly anaerobic culture
-
native enzyme is a three-step procedure
-
of anaerobically grown cells
-
partial purification under anaerobic conditions
-
purification aerobically at 4C
-
purification under anaerobic conditions
recombinant His6-tagged enzyme and Strep-tagged enzyme from Escherichia coli strain BL21(DE3) with or without bound calcium by metal affinity and streptavidin chromatography, respectively, followed by gel filtration
recombinant metal-free apo protein
-
two forms a purple and a blue form
-