Information on EC 1.7.2.4 - nitrous-oxide reductase

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The expected taxonomic range for this enzyme is: Bacteria, Archaea

EC NUMBER
COMMENTARY
1.7.2.4
-
RECOMMENDED NAME
GeneOntology No.
nitrous-oxide reductase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
nitrogen + H2O + 2 ferricytochrome c = nitrous oxide + 2 ferrocytochrome c + 2 H+
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
oxidation
-
-
-
-
redox reaction
-
-
-
-
reduction
-
-
-
-
PATHWAY
KEGG Link
MetaCyc Link
Microbial metabolism in diverse environments
-
nitrate reduction I (denitrification)
-
nitrate reduction VII (denitrification)
-
nitrifier denitrification
-
Nitrogen metabolism
-
SYSTEMATIC NAME
IUBMB Comments
nitrogen:cytochrome c oxidoreductase (N2O-forming)
The reaction is observed only in the direction of nitrous oxide reduction. Contains the mixed-valent dinuclear CuA species at the electron entry site of the enzyme, and the tetranuclear Cu-Z centre in the active site. In Paracoccus pantotrophus, the electron donor is cytochrome c552.
SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
HdN2OR
Hyphomicrobium denitrificans A3151
-
-
-
multicopper oxidase
-
-
N(2)OR
-
-
-
-
N2O reductase
-
-
-
-
N2O reductase
P94127
-
N2O reductase
-
-
N2O reductase
Bradyrhizobium japonicum USDA110
-
-
-
N2O reductase
-
-
N2O reductase
Ensifer adhaerens SN611
-
-
-
N2O reductase
-
-
N2O reductase
-
-
N2O reductase
Pseudomonas denitrificans CCUG 2519
-
-
-
N2O reductase
Q9F0W4
-
N2O reductase
Pseudomonas fluorescens C7R12
Q9F0W4
-
-
N2O reductase
-
-
N2O reductase
Sinorhizobium meliloti 50
-
-
-
N2OR
Achromobacter cycloclastes IAM1013
P94127
-
-
nitrous oxide reductase
-
-
-
-
nitrous oxide reductase
-
-
nitrous oxide reductase
P94127
-
nitrous oxide reductase
Achromobacter cycloclastes IAM1013
P94127
-
-
nitrous oxide reductase
-
-
nitrous oxide reductase
Bradyrhizobium japonicum USDA110
-
-
-
nitrous oxide reductase
Q19Q69
-
nitrous oxide reductase
Q51705
-
nitrous oxide reductase
-
-
nitrous oxide reductase
-
-
nitrous oxide reductase
-
-
NosZ
Q89XJ6
catalytic subunit of nitrous oxide reductase
NosZ
Bradyrhizobium japonicum USDA110
-
-
-
NosZ
Ensifer adhaerens SN611
-
-
-
NosZ
Q51705
gene name
NosZ
Pseudomonas denitrificans CCUG 2519
-
-
-
NosZ
Pseudomonas fluorescens C7R12
Q9F0W4
-
-
NosZ
-
-
-
NosZ
Pseudomonas stutzeri ATCC 14405
P19573
-
-
NosZ
Sinorhizobium meliloti 50
-
-
-
CAS REGISTRY NUMBER
COMMENTARY
55576-44-8
-
ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
strain ATCC 21921
-
-
Manually annotated by BRENDA team
strain IAM1013
UniProt
Manually annotated by BRENDA team
Achromobacter cycloclastes 1013
strain 1013
UniProt
Manually annotated by BRENDA team
Achromobacter cycloclastes IAM1013
strain IAM1013
UniProt
Manually annotated by BRENDA team
NCIMB 11015, X-ray scattering data, 20 A resolution
-
-
Manually annotated by BRENDA team
strain ATCC 8750
-
-
Manually annotated by BRENDA team
strain 562 and strain USDA110
-
-
Manually annotated by BRENDA team
strain USDA110
-
-
Manually annotated by BRENDA team
Bradyrhizobium japonicum USDA110
strain USDA110
-
-
Manually annotated by BRENDA team
strain DSM 15892T
-
-
Manually annotated by BRENDA team
strain SN611
-
-
Manually annotated by BRENDA team
Ensifer adhaerens SN611
strain SN611
-
-
Manually annotated by BRENDA team
strain DSM 15936T
-
-
Manually annotated by BRENDA team
strain DSM 1869
-
-
Manually annotated by BRENDA team
Hyphomicrobium denitrificans A3151
A3151
-
-
Manually annotated by BRENDA team
Marinobacter hydrocarbonoclasticus 617
strain 617
-
-
Manually annotated by BRENDA team
formerly Thiosphaera pantotropha
-
-
Manually annotated by BRENDA team
formerly Thiosphaera pantotropha; LMD 82.5
-
-
Manually annotated by BRENDA team
Paracoccus pantotrophus LMD 82.5
LMD 82.5
-
-
Manually annotated by BRENDA team
Pseudomonas aeruginosa P2
strain P2
-
-
Manually annotated by BRENDA team
strain CCUG 2519
-
-
Manually annotated by BRENDA team
Pseudomonas denitrificans CCUG 2519
strain CCUG 2519
-
-
Manually annotated by BRENDA team
precursor; strain C7R12
SwissProt
Manually annotated by BRENDA team
Pseudomonas fluorescens C7R12
precursor; strain C7R12
SwissProt
Manually annotated by BRENDA team
strain ED3
-
-
Manually annotated by BRENDA team
strain G59
-
-
Manually annotated by BRENDA team
strain ED3
-
-
Manually annotated by BRENDA team
Pseudomonas sp. G59
strain G59
-
-
Manually annotated by BRENDA team
wild-type, C622D-mutant laccking copper in the CuA-site and N2OR V a mutant which lacks CuZ
-
-
Manually annotated by BRENDA team
Pseudomonas stutzeri ATCC 14405
-
UniProt
Manually annotated by BRENDA team
strain 8A55
-
-
Manually annotated by BRENDA team
Rhizobium sp. 8A55
strain 8A55
-
-
Manually annotated by BRENDA team
strains N22, Kb1
-
-
Manually annotated by BRENDA team
f. sp. denitrificans IL106
-
-
Manually annotated by BRENDA team
strain 2.4.1
-
-
Manually annotated by BRENDA team
Rhodopseudomonas palustris PW5
strain PW5
-
-
Manually annotated by BRENDA team
Rhodospirillum rubrum S1
stain S1
-
-
Manually annotated by BRENDA team
strain 1021; strain 50
-
-
Manually annotated by BRENDA team
Sinorhizobium meliloti 50
strain 50
-
-
Manually annotated by BRENDA team
ATCC 29591
-
-
Manually annotated by BRENDA team
under anaerobic conditions cytochrome c oxidase activity is observed
-
-
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
N2O + 2 Fe2+ + 2 H+
N2 + H2O + 2 Fe3+
show the reaction diagram
-
-
-
-
?
N2O + H2O + benzyl viologen cation radical
N2 + reduced benzyl viologen
show the reaction diagram
Pseudomonas aeruginosa, Pseudomonas aeruginosa P2
-
-
-
-
?
N2O + reduced acceptor
N2 + H2O + acceptor
show the reaction diagram
Q51705
-
-
-
?
N2O + reduced acceptor
N2 + H2O + acceptor
show the reaction diagram
-
-
-
-
?
N2O + reduced acceptor
N2 + H2O + acceptor
show the reaction diagram
-
-
-
-
?
N2O + reduced acceptor
N2 + H2O + acceptor
show the reaction diagram
-
-
-
-
?
N2O + reduced acceptor
N2 + H2O + acceptor
show the reaction diagram
-
-
-
-
?
N2O + reduced acceptor
N2 + H2O + acceptor
show the reaction diagram
-
-
-
-
?
N2O + reduced acceptor
N2 + H2O + acceptor
show the reaction diagram
-
-
-
-
?
N2O + reduced acceptor
N2 + H2O + acceptor
show the reaction diagram
-
-
-
-
?
N2O + reduced acceptor
N2 + H2O + acceptor
show the reaction diagram
-
-
-
-
?
N2O + reduced acceptor
N2 + H2O + acceptor
show the reaction diagram
P94127
-
-
-
?
N2O + reduced acceptor
N2 + H2O + acceptor
show the reaction diagram
-
-
-
-
?
N2O + reduced acceptor
N2 + H2O + acceptor
show the reaction diagram
-, Q89XJ6
-
-
-
?
N2O + reduced acceptor
N2 + H2O + acceptor
show the reaction diagram
-
-
-
-
?
N2O + reduced acceptor
N2 + H2O + acceptor
show the reaction diagram
-
-
-
-
?
N2O + reduced acceptor
N2 + H2O + acceptor
show the reaction diagram
Q9F0W4
-
-
-
?
N2O + reduced acceptor
N2 + H2O + acceptor
show the reaction diagram
-
-
-
-
?
N2O + reduced acceptor
N2 + H2O + acceptor
show the reaction diagram
Pseudomonas denitrificans CCUG 2519
-
-
-
-
?
N2O + reduced acceptor
N2 + H2O + acceptor
show the reaction diagram
Pseudomonas fluorescens C7R12
Q9F0W4
-
-
-
?
N2O + reduced acceptor
N2 + H2O + acceptor
show the reaction diagram
Sinorhizobium meliloti 50
-
-
-
-
?
N2O + reduced acceptor
N2 + H2O + acceptor
show the reaction diagram
Hyphomicrobium denitrificans A3151
-
-
-
-
?
N2O + reduced acceptor
N2 + H2O + acceptor
show the reaction diagram
Bradyrhizobium japonicum USDA110
-
-
-
-
?
N2O + reduced acceptor
N2 + H2O + acceptor
show the reaction diagram
Ensifer adhaerens SN611
-
-
-
-
?
N2O + reduced benzyl viologen
N2 + H2O + benzyl viologen
show the reaction diagram
-
-
-
-
-
N2O + reduced benzyl viologen
N2 + H2O + benzyl viologen
show the reaction diagram
-
-
-
-
?
N2O + reduced benzyl viologen
N2 + H2O + benzyl viologen
show the reaction diagram
Hyphomicrobium denitrificans, Hyphomicrobium denitrificans A3151
-
-
-
-
?
N2O + reduced benzyl viologen
N2 + oxidized benzyl viologen
show the reaction diagram
-
-
-
-
?
N2O + reduced cytochrome c
N2 + H2O + cytochrome c
show the reaction diagram
-
-
-
-
?
N2O + reduced cytochrome c550
N2 + H2O + cytochrome c550
show the reaction diagram
Q51705
-
-
-
?
N2O + reduced cytochrome c550
N2 + H2O + cytochrome c550
show the reaction diagram
-
-
-
-
?
N2O + reduced methyl viologen
N2 + H2O + methyl viologen
show the reaction diagram
-
-
-
-
?
N2O + reduced methyl viologen
N2 + H2O + methyl viologen
show the reaction diagram
-
-
-
-
?
N2O + reduced methyl viologen
N2 + H2O + oxidized methyl viologen
show the reaction diagram
-
-
-
-
?
N2O + reduced methyl viologen
N2 + oxidized methyl viologen
show the reaction diagram
Pseudomonas stutzeri, Pseudomonas stutzeri ATCC 14405
P19573
-
-
-
?
N2O + reduced pseudoazurin
N2 + H2O + pseudoazurin
show the reaction diagram
Q51705
-
-
-
?
N2O + reduced pseudoazurin
N2 + H2O + pseudoazurin
show the reaction diagram
-
-
-
-
?
N2O + reduced pseudoazurin
N2 + oxidized pseudoazurin
show the reaction diagram
-
blue copper electron-transfer protein pseudoazurin is capable of mediating electron transfer to the nitrous oxide reductase. Pseudoazurin binds near copper site CuA, with parameters consistent with the formation of a transient, weakly-bound complex. Pseudoazurin a strong candidate for the physiological electron donor to th enzyme
-
-
?
nitrogen + H2O + acceptor
nitrous oxide + reduced acceptor
show the reaction diagram
-
-
-
-
?
nitrogen + H2O + acceptor
nitrous oxide + reduced acceptor
show the reaction diagram
P94127
-, under some conditions, electron transfer may be rate limiting in N2O reduction. The transition state is stabilized by H-bonding interactions between the active site and an N2O-derived intermediate bound to the catalytic CuZ cluster
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
-
-
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
-
-
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
-
-
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
-
-
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
-
-
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
-
-
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
-
-
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
-
-
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
-
-
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
-
-
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
-
-
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
-
-
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
-
-
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
-
-
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
-
-
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
-
-
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
-
methyl viologen as electron donor
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
-
methyl viologen as electron donor
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
-
methyl viologen as electron donor
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
-
methyl viologen as electron donor, activity after incubation with ferricyanide, ascorbate or dithionite
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
-
photochemically-reduced benzyl viologen as electron donor
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
-
photochemically-reduced benzyl viologen as electron donor
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
-
photochemically-reduced benzyl viologen as electron donor
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
-
photochemically-reduced benzyl viologen as electron donor
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
-
photochemically-reduced benzyl viologen as electron donor
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
-
photochemically reduced benzyl viologen and 3,3',5,5'-tetramethylbenzidine as electron donor
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
-
a mixture of N,N,N',N'-tetramethyl-p-phenylenediamine and ascorbate as electron donor
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
-
dithionite-reduced methyl viologen as electron donor
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
-
dithionite-reduced methyl viologen as electron donor
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
-
dithionite-reduced benzyl viologen as electron donor
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
Paracoccus pantotrophus LMD 82.5
-
-, dithionite-reduced methyl viologen as electron donor
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
Rhodospirillum rubrum S1
-
-
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
Rhizobium sp. 8A55
-
-
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
Pseudomonas sp. G59
-
-
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
Rhodopseudomonas palustris PW5
-
-
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
Marinobacter hydrocarbonoclasticus 617
-
-
-
-
?
nitrous oxide + reduced methyl viologen
nitrogen + H2O + methyl viologen
show the reaction diagram
Achromobacter cycloclastes, Achromobacter cycloclastes IAM1013
P94127
-
-
-
?
additional information
?
-
-
enzyme shows Cu+/Fe2+ oxidation kinetics that follow the Michaelis-Menten model, with 2fold to 10fold higher efficiencies for Cu+ and Fe2+ as compared with the tested aromatic compounds
-
-
-
additional information
?
-
-
the purple form of enzyme, in which the copper centre is in the oxidized [2Cu2+:2Cu+] redox state, is redox active, although it is still catalytically non-competent, as its specific activity is lower than that of the activated fully reduced enzyme and comparable with that of the enzyme with the copper centre in either the [1Cu2+:3Cu+] redox state or in the redox inactive state
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
N2O + reduced acceptor
N2 + H2O + acceptor
show the reaction diagram
Q51705
-
-
-
?
N2O + reduced acceptor
N2 + H2O + acceptor
show the reaction diagram
-
-
-
-
?
N2O + reduced acceptor
N2 + H2O + acceptor
show the reaction diagram
-
-
-
-
?
N2O + reduced acceptor
N2 + H2O + acceptor
show the reaction diagram
-
-
-
-
?
N2O + reduced acceptor
N2 + H2O + acceptor
show the reaction diagram
-, Q89XJ6
-
-
-
?
N2O + reduced acceptor
N2 + H2O + acceptor
show the reaction diagram
Hyphomicrobium denitrificans A3151
-
-
-
-
?
N2O + reduced pseudoazurin
N2 + oxidized pseudoazurin
show the reaction diagram
-
blue copper electron-transfer protein pseudoazurin is capable of mediating electron transfer to the nitrous oxide reductase. Pseudoazurin binds near copper site CuA, with parameters consistent with the formation of a transient, weakly-bound complex. Pseudoazurin a strong candidate for the physiological electron donor to th enzyme
-
-
?
nitrogen + H2O + acceptor
nitrous oxide + reduced acceptor
show the reaction diagram
P94127
-
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
-
-
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
-
-
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
-
-
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
-
-
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
-
-
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
-
-
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
-
-
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
-
-
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
-
-
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
-
-
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
-
-
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
-
-
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
-
-
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
-
-
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
-
-
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
-
-
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
Paracoccus pantotrophus LMD 82.5
-
-
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
Rhodospirillum rubrum S1
-
-
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
Rhizobium sp. 8A55
-
-
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
Pseudomonas sp. G59
-
-
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
Rhodopseudomonas palustris PW5
-
-
-
-
?
nitrous oxide + reduced acceptor
nitrogen + H2O + acceptor
show the reaction diagram
Marinobacter hydrocarbonoclasticus 617
-
-
-
-
?
nitrous oxide + reduced methyl viologen
nitrogen + H2O + methyl viologen
show the reaction diagram
Achromobacter cycloclastes, Achromobacter cycloclastes IAM1013
P94127
-
-
-
?
COFACTOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
cupredoxin
-
a copper enzyme with cupredoxin containing blue T1 copper and red T2 copper. Blue and red copper centers form initially before they are pH-dependently transformed into purple CuA center, lower pH resulting in fewer trapped T1 and T2 coppers and faster transition. Structure overview
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Ca2+
-
two calcium sites located at the intermonomeric surface
Ca2+
-
1.2 atoms per subunit
copper
-
the fully reduced all-Cu(I) state of CuZ is the catalytically relevant redox state of N2OR
copper
-
form A and B contain 9.0 and 8.2 Cu atoms per dimer, respectively
copper
-
multicopper enzyme, 2 copper centers per subunit, CuA and CuZ
copper
-
N2OR contains two copper centers, CuA, a binuclear mixed-valence center and CuZ, a tetranuclear sulfide-bridged copper cluster
copper
-
3.2 mol of copper per mol of enzyme, in the as-isolated form. Presence of 0.1 mM enhances enzymic activity by 2fold
Cu
-
one dinuclear centre CuA and a copper cluster CuZ in which four copper ions are litigated by seven histidine imidazoles and a bridging inorganic sufide
Cu
-
CuA can exist in two oxidation forms [Cu1.5+ - Cu1.5+] and [Cu1+ - Cu1+]; one dinuclear centre CuA and a copper cluster CuZ in which four copper ions are litigated by seven histidine imidazoles and a bridging inorganic sufide
Cu
-
wild-type 10.5 atoms per protein dimer
Cu
-
wild-type 9.9 atoms per protein dimer
Cu
-
six copper atoms per monomer arranged in two centers named CuA and CuZ, 10.7 atoms per dimer
Cu
-
contains 7.1 atoms per dimer
Cu
-
7.0 atoms per dimer, needs 0.001 mM Cu2+ for full activity
Cu
-
CuA is electron transfer center, CuZ is the catalytic center
Cu
-
wild-type 7.2 atoms per protein dimer
Cu
-
4 atoms per subunit
Cu
-
2.9 atoms per subunit
Cu
-
4.0 atoms per subunit
Cu
-
4 atoms per mol
Cu
-
8 atoms per mol enzyme
Cu
-
A monomer carries 2 copper centers: CuA, which is a binuclear mixed-valence center similar to the CuA of cytochrome oxidases,4 and CuZ, a tetranuclear sulfidebridged copper cluster.
Cu
-
4 Cu atoms/subunit
Cu
-
Cu is bound by apo NosL, a coexpressed protein which is necessary for the assembling process of nitrous oxide reductase
Cu
-
N2OR contains two metal centers: a binuclear copper center, CuA, that serves to receive electrons from soluble donors, and a tetranuclear copper-sulfide center, CuZ, at the active site.
Cu
-
The two copper atoms (CuI and CuIV) at the ligand-binding site of the cluster play a crucial role in the enzymatic function, as these atoms are directly involved in bridged N2O binding.
Cu
P94127
mu4-sulfide-bridged tetranuclear CuZ cluster
Cu2+
-
a copper enzyme, conformational changes of the CuA center may affect electron transfer from the physiological electron donor and the electron-transfer rate from CuA to CuZ, overview
Cu2+
-
a copper enzyme, two copper centers, CuA and CuB, per monomer, binding structure, detailed analysis of the pH effect at the CuZ site, e.g. using spectroscopic and computational methods, overview
Cu2+
Q19Q69
a copper enzyme, two copper centers, CuA and CuB, binding structure, detailed analysis of the pH effect at the CuZ site, e.g. using spectroscopic and computational methods, overview
Cu2+
P94127
the enzyme contains average 4.5 Cu and 1.2 S per monomer
Cu2+
-
the copper enzyme contains an unusual mixed valence copper, Cu(I)/Cu(II), dimer centre, the primary paramagnetic species is the CuA, the catalytic CuZ centre being primarily in a diamagnetic oxidized form. Coherent Raman detected electron spin resonance spectroscopy structure analysis of the CuA site in nitrous oxide reductase, overview
Cu2+
-
a copper enzyme with cupredoxin containing blue T1 copper and red T2 copper. Blue and red copper centers form initially before they are pH-dependently transformed into purple CuA center, lower pH resulting in fewer trapped T1 and T2 coppers and faster transition
Fe
-
1 atom per mol
Ni
-
0.76 atoms per mol
Zn
-
2 atoms per mol
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
2-mercaptoethanol
-
53% after 30 min at 0.5 mM
Acetylene
-
at 10 mM 80% inhibition
Acetylene
-
at 4 kPa; together with sulfide no inhibition
Acetylene
-
at 10% complete inhibition
EDTA
-
complete inhibition at 0.1 mM
EDTA
-
11% at 0.5 mM after 30 min
EGTA
-
36% at 0.5 mM after 30 min
N,N,N',N'-tetramethyl-p-phenylenediamine
-
76% inhibition at 0.1 mM
Na2S
-
temporarily inhibition; together with sulfide no inhibition
Ni2+
-
50% inhibition at 0.1 mM
O2
-
inactivation is faster in the crude extract than solutions of the purified enzyme, acceleration takes place if the enzyme is turning over
O2
-
turnover dependent inactivation
O2
-
complete inactivation at 0.25 mg/l, no activity restorage after return to anaerobic conditions
O2
-
unpurified enzyme is rapid inactivated at air
O2
-
inactivation with O2 is largely reversible by exposure to benzyl viologen cation radical at room temperature for several hours
O2
-
50% inactivation after 30 min, 80% regained activity after incubation with reduced benzyl viologen
rotenone
-
88% inhibition at 0.025 mM
S2O42-
-
irreversible formation of inactive blue form of the enzyme
S2O42-
-
irreversible formation of inactive blue form of the enzyme
sodium iodide
P94127
0.2 M
Zn2+
-
complete inhibition at 0.1 mM
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
NO2-
-
increases the synthesis of N2O-reductase
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.004
-
benzyl viologen
-
-
0.0043
-
benzyl viologen cation radical
-
pH 7.1, 25C
0.0024
-
N2O
-
pH 7.1, 25C
0.0191
-
N2O
-
pH 7.1, temperature not specified in the publication
0.032
-
N2O
-
pH 7.6, 25C
0.0009
-
reduced benzyl viologen
-
-
0.0138
-
reduced benzyl viologen
-
pH 7.1, temperature not specified in the publication
0.00095
-
reduced methyl viologen
-
-
0.0288
-
reduced pseudoazurin
-
pH 7.1, temperature not specified in the publication
-
0.033
-
Fe2+
-
pH 7.6, 25C
-
additional information
-
additional information
-
steady-state kinetics, overview
-
additional information
-
additional information
P94127
steady-state kinetics
-
TURNOVER NUMBER [1/s]
TURNOVER NUMBER MAXIMUM[1/s]
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
8
-
N2O
-
pH 7.6, 25C
162.9
-
reduced benzyl viologen
-
pH 7.1, temperature not specified in the publication
89.3
-
reduced pseudoazurin
-
pH 7.1, temperature not specified in the publication
-
kcat/KM VALUE [1/mMs-1]
kcat/KM VALUE [1/mMs-1] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
242
-
Fe2+
-
pH 7.6, 25C
0
250
-
N2O
-
pH 7.6, 25C
14203
Ki VALUE [mM]
Ki VALUE [mM] Maximum
INHIBITOR
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.028
-
Acetylene
-
non competitive
0.035
-
Acetylene
-
-
0.0035
-
CO
-
non competitive
0.000045
-
KCN
-
non competitive
0.00035
-
NaN3
-
non competitive
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
0.4
-
-
fully oxidized enzyme isolated from aerobic growth conditions, pH 7.6, temperature not specified in the publication
1
-
-
ascorbate-reduced phenanzine methosulphate electrode assay
1.1
-
P94127
assayed in the standard dithionite methyl viologen assay
1.1
-
-
fully oxidized enzyme isolated from aanerobic growth conditions, pH 7.6, temperature not specified in the publication
1.6
-
-
anaerobic form pre-reduced with ascorbate
1.97
-
-
wild type enzyme
2.6
-
-
anaerobic form pre-oxidized with ferricyanide
2.8
-
-
anaerobic form pre-reduced with dithionite
3.3
-
-
photoreduced methyl viologen spectrophotometric assay
3.52
-
-
single mutant nirX
3.63
-
-
double mutant nosXnirX
3.7
-
-
dithionite-reduced methyl viologen electrode assay
5.5
-
-
wild-type
8.5
-
-
aerobic form pre-oxidized with ferricyanide
8.8
-
-
aerobic form pre-reduced with dithionite
23
-
-
blue-coloured form
27
-
-
pH 7.1, 25C
29
-
-
HdN2OR form A, i.e. reduced form
44
-
-
aerobic form
45
-
-
HdN2OR form A, i.e. oxidized form
55
-
-
purple coloured form
61.7
-
-
after preincubation in 2 mM reduced benzyl viologen
78
-
-
anaerobic form
88
-
-
fully reduced enzyme isolated from aerobic growth conditions, pH 7.6, temperature not specified in the publication
92.4
-
-
fully reduced enzyme isolated from aerobic growth conditions, pH 7.6, temperature not specified in the publication
122
-
-
anaerobic form
124
-
P94127
purified enzyme, pH 8.0, 25C
453
-
P19573
enzyme purified from transgenic Nicotiana tabacum, pH 7.1, 37C
additional information
-
-
Vmax N2O 0.2 - 0.5 micromol/min/mg, pH 7.1, 25C
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7.1
8
P94127
assay at
8
-
-
above pH 8 with reduced methyl viologen
8
-
-
in presence of N,N,N',N'-tetramethyl-p-phenylenediamine and ascorbate
8.5
9
-
with reduced benzyl viologen as electron donor
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
5.7
10.6
P94127
perturbations of the protein conformation induced by pH variations, although the principal secondary structure elements are largely unaltered
5.7
9.4
-
pH profile with two maxima, high complexity of pH dependence, overview
6
9.2
-
half maximum activity values at pH 6 and pH 9.2
7
9.5
-
approx. 50% of maximal activity at pH 7.5, approx. 75% of maximal activity at pH 9.5
9
10
-
activity is maximal after incubation at high pH values
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
25
-
P94127
assay at
TEMPERATURE RANGE
TEMPERATURE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
40
85
-
activity is maximal after exposure to high temperatures
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
Marinobacter hydrocarbonoclasticus 617
-
soluble
-
-
Manually annotated by BRENDA team
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
49600
-
-
gel fitlration
85000
-
-
gel filtration
89000
-
-
gel filtration
95000
-
-
gel filtration
105000
-
-
gel filtration
105000
-
-
gel filtration
115000
-
-
gel filtration
118000
-
-
gel filtration
120000
-
-
gel filtration
120000
-
-
gel filtration
120000
-
-
HPLC gel filtration
130000
-
-
non denaturating PAGE
134000
-
-
gel filtration
144000
-
-
gel filtration
160000
-
-
gel filtration
162000
-
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-, Q89XJ6
x * 71670, deduced from nucleotide sequence
dimer
-
2 * 65000, SDS-PAGE
dimer
-
2 * 73000, SDS-PAGE
dimer
Hyphomicrobium denitrificans A3151
-
2 * 65000, SDS-PAGE
-
dimer
Pseudomonas aeruginosa P2
-
2 * 73000, SDS-PAGE
-
homodimer
-
2 * ?, each monomer is composed of two domains, a C-terminal cupredoxin domain carrying a dinuclear electron entry site CuA, an N-terminal seven-bladed propeller domain with the active center CuZ
homodimer
-
2 * 65000, SDS-PAGE; 2 * 65373, ESI mass spectrometry
homodimer
-
2 * ?, each monomer is composed of two domains, a C-terminal cupredoxin domain carrying a dinuclear electron entry site CuA, an N-terminal seven-bladed propeller domain with the active center CuZ
homodimer
-
2 * 66180, MALDI-TOF
homodimer
-
2 * 66871, ESI mass spectrometry
homodimer
-
2 * 66180, MALDI-TOF; 2 * 67000, SDS-PAGE
homodimer
-
2 * 73000, SDS-PAGE
homodimer
-
2 * 66281, ESI mass spectrometry; 2 * 73000, SDS-PAGE
homodimer
-
2 * 88000, SDS-PAGE
homodimer
-
2 * 70000, SDS-PAGE
homodimer
-
2 * 74000, SDS-PAGE
homodimer
-
alpha2, 2 * 62000, SDS-PAGE
homodimer
-
2 * 65000, X-ray crystallography
homodimer
P94127
x-ray crystallography
homodimer
Marinobacter hydrocarbonoclasticus 617
-
2 * 65000, SDS-PAGE; 2 * 65373, ESI mass spectrometry; 2 * ?, each monomer is composed of two domains, a C-terminal cupredoxin domain carrying a dinuclear electron entry site CuA, an N-terminal seven-bladed propeller domain with the active center CuZ
-
homodimer
Paracoccus pantotrophus LMD 82.5
-
2 * 66281, ESI mass spectrometry; 2 * 73000, SDS-PAGE
-
monomer
-
1 * 72000, SDS-PAGE
monomer
-
1 * 52900, calculated, 1 * 52000, SDS-PAGE
additional information
P94127
perturbations of the protein conformation induced by pH variations, although the principal secondary structure elements are largely unaltered
additional information
-
the enzyme contains an unusual mixed valence copper, Cu(I)/Cu(II), dimer centre, coherent Raman detected electron spin resonance spectroscopy structure analysis of the CuA site in nitrous oxide reductase, overview
additional information
Achromobacter cycloclastes IAM1013
-
perturbations of the protein conformation induced by pH variations, although the principal secondary structure elements are largely unaltered
-
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
-
the purple form of enzyme, in which the copper centre is in the oxidized [2Cu2+:2Cu+] redox state, is redox active, although it is still catalytically non-competent, as its specific activity is lower than that of the activated fully reduced enzyme and comparable with that of the enzyme with the copper centre in either the [1Cu2+:3Cu+] redox state or in the redox inactive state
Crystallization/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
hanging drop vapour diffusion method
P94127
2.4 A resolution, preliminary study
-
1.6 A resolution
-
building of two models of the active site reveals two distinct mechanisms. In the first model, N2O binds to the fully reduced tetranuclear Cu4S core in a bent my-(1,3)-O,N bridging fashion between the CuI and CuIV centres and subsequently extrudes N2 while generating the corresponding bridged my-oxo species. In the second model, substrate N2O binds loosely to one of the coppers of the tetranuclear Cu4S core in a terminal fashion, i.e., using only the oxygen atom. Loss of N2 generates the same my-oxo copper core. The free energies of activation predicted for these two alternative pathways are close to one another and do not provide decisive support for one over the other
-
modeling of structure. The residues contributing to the semiocclusion of the T1 copper site are Trp355, Met389, and Met297. There is a negatively charged residue in the neighborhood of the T1 site, Glu296
-
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
25
-
-
85% inactivation after 20 h
70
-
-
most of activity lost
85
-
-
half-life 5.5 h
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
turnover dependent activation, nitrite and fluoride accelerate this process
-
an enzyme concentration of at least 0.01 mg per ml is necessary to avoid loss of activity due to dilution
-
turnover dependent inactivation, promoted by zinc ions at 0.01 mM
-
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
4C, retained most of its activity after aerobic storage for 2 months without any additives
-
4C, under aerobic conditions, 100 h stable
-
4C, under aerobic conditions, 5 mM EDTA, 3 h stable
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
anaerobical purification of N2OR under an Ar atmosphere by three different steps of anion exchange chromatography, followed by cytochrome c affinity chromatography and ultrafiltration
P94127
DEAE cellulose column chromatography, hydroxyapatite column chromatography, Mono Q10 anion-exchange column chromatography, and Sephadex S-200 gel filtration
P94127
DEAE-FF ion exchange chromatography and Resource Q column chromatography
-
purification under anaerobic conditions
-
ammonium sulfate, DEAE-Sephadex A-50, Sephacryl S-200
-
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
-
two forms a purple and a blue form
-
HiTrap Q Sepharose anion exchange chromatography
-
partial purification under anaerobic conditions
-
purification under anaerobic conditions
-
recombinant metal-free apo protein
-
of anaerobically grown cells
-
a pink form is isolated when the purification is done aerobically, a purple form when the purification is under anaerobic conditions
-
purification aerobically at 4C
-
purification under anaerobic conditions
-
purification under anaerobic conditions
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
expressed in Escherichia coli
-
the gene encoding the enzyme in the nos cluster is cloned from a genomic library
P94127
expression as metal-free apo protein
-
cloning of the nos gene cluster
-
expression in Escherichia coli HB101, no activity
-
expression in Nicotiana tabacum
P19573
expression in Escherichia coli
-
ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
C165G
-
Cys165 is not available for Cu cooordination; retaines catalytic activity
C622D
-
no activity, distorted CuA-centre
E296Q
-
mutation near T1 copper site, similar biochemical and spectroscopic properties to those of the wild type
M297A
-
mutation near T1 copper site, similar biochemical and spectroscopic properties to those of the wild type
M389A
-
mutation near T1 copper site, similar biochemical and spectroscopic properties to those of the wild type
W355A
-
mutation near T1 copper site, similar biochemical and spectroscopic properties to those of the wild type
additional information
-
deletion mutant USDA110DELTAnosZ shows no N2O reductase activity
additional information
Bradyrhizobium japonicum USDA110
-
deletion mutant USDA110DELTAnosZ shows no N2O reductase activity
-
additional information
-
strains with a nosXnirX double mutation or nirX single mutation show loss of N2O respiration
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
degradation
-
plays a critical enviromental role in preventing release into the atmosphere of the potent greenhouse gas nitrous oxide
agriculture
P19573
expression of both the senzyme-coding gene nosZ and the mega-cassette of five coding sequences nosFLZDY in Nicotiana tabacum leads to active recombinant N2OR. Extracts from both types of transgenic plants exhibit N2O-reducing activity. The single-gene strategy produces higher reductase capability than the whole-operon approach. Bacterial nitrous oxide reductase expressed in plants could convert N2O into inert N2 without involvement of other Nos proteins
degradation
-
plays a critical enviromental role in preventing release into the atmosphere of the potent greenhouse gas nitrous oxide
agriculture
Pseudomonas stutzeri ATCC 14405
-
expression of both the senzyme-coding gene nosZ and the mega-cassette of five coding sequences nosFLZDY in Nicotiana tabacum leads to active recombinant N2OR. Extracts from both types of transgenic plants exhibit N2O-reducing activity. The single-gene strategy produces higher reductase capability than the whole-operon approach. Bacterial nitrous oxide reductase expressed in plants could convert N2O into inert N2 without involvement of other Nos proteins
-