Information on EC 1.14.14.10 - nitrilotriacetate monooxygenase

Word Map on EC 1.14.14.10
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Select one or more organisms in this record:
Show additional data
Do not include text mining results
Include (text mining) results (more...)
Include results (AMENDA + additional results, but less precise; more...)


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

EC NUMBER
COMMENTARY hide
1.14.14.10
-
RECOMMENDED NAME
GeneOntology No.
nitrilotriacetate monooxygenase
-
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
nitrilotriacetate + FMNH2 + H+ + O2 = iminodiacetate + glyoxylate + FMN + H2O
show the reaction diagram
-
-
-
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oxidation
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
nitrilotriacetate degradation
-
-
SYSTEMATIC NAME
IUBMB Comments
nitrilotriacetate,FMNH2:oxygen oxidoreductase (glyoxylate-forming)
Requires Mg2+. The enzyme from Aminobacter aminovorans (previously Chelatobacter heintzii) is part of a two component system that also includes EC 1.5.1.42 (FMN reductase), which provides reduced flavin mononucleotide for this enzyme.
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
Chelativorans sp.
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
EDTA + O2 + FMNH2 + H+
ethylenediaminetriacetate + glyoxylate + H2O + FMN
show the reaction diagram
Chelativorans sp.
-
-
-
-
?
nitrilotriacetate + FMNH2 + H+ + O2
iminodiacetate + glyoxylate + FMN + H2O
show the reaction diagram
nitrilotriacetate + O2 + FMNH2 + H+
iminodiacetate + glyoxylate + H2O + FMN
show the reaction diagram
Chelativorans sp.
-
-
-
-
?
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
nitrilotriacetate + FMNH2 + H+ + O2
iminodiacetate + glyoxylate + FMN + H2O
show the reaction diagram
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
FMN
-
omission of FMN or replacement of FMN by FAD results in a residual activity of 2% when component B is not resaturated with FMN
FMNH2
NADH
-
NADPH cannot replace NADH
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Al3+
-
nitrilotriacetate is only a substrate when complexed with cations such as Mg2+, Al3+, Cu2+, Ni2+, Zn2+, Fe2+ or Co2+
Cu2+
-
nitrilotriacetate is only a substrate when complexed with cations such as Mg2+, Al3+, Cu2+, Ni2+, Zn2+, Fe2+ or Co2+
Fe2+
-
nitrilotriacetate is only a substrate when complexed with cations such as Mg2+, Al3+, Cu2+, Ni2+, Zn2+, Fe2+ or Co2+
Mn2+
-
Mn2+ ions are able to replace Mg2+ but lead to a higher uncoupled NADH oxidation and enzyme activity is reduced to 70% of that with MgCl2
Ni2+
-
nitrilotriacetate is only a substrate when complexed with cations such as Mg2+, Al3+, Cu2+, Ni2+, Zn2+, Fe2+ or Co2+
Zn2+
-
nitrilotriacetate is only a substrate when complexed with cations such as Mg2+, Al3+, Cu2+, Ni2+, Zn2+, Fe2+ or Co2+
additional information
-
no nitrilotriacetate consumption is observed with Ca2+,Fe2+, Fe3+, Zn2+, Cu2+, or Ni2+; the enzyme contains less than 0.15 atom of Fe per mol of protein, indicating that neither Fe-sulfur clusters nor cytochromes are present
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
5,5'-dithiobis(2-nitrobenzoate)
-
67% residual activity at 1 mM
Ca2+
-
5% residual activity at 10 mM
EDTA
-
3% residual activity at 10 mM
FAD
-
24% residual activity at 0.1 mM
NaCl
-
54% residual activity at 250 mM
Ni2+
-
ionic or uncomplexed Ni2+ (2 mM) is inhibitory to the enzyme
p-hydroxymercuribenzoate
-
12% residual activity at 1 mM
Zn2+
-
ionic or uncomplexed Zn2+ (2 mM) is inhibitory to the enzyme
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
component A of NTA monooxygenase
-
the ability of component B of NTA monooxygenase to catalyze the oxidation of nitrilotriacetate to iminodiacetate and glyoxylate is completely dependent on the presence of component A
-
FMN
-
NTA-Mo activity increases by a factor of 4 when 0.003 mM FMN is included in the assay mixture, there is 0.4 mol of FMN per mol of purified component B
additional information
-
the activity is increased neither by additions of 20% (w/v) polyethylene glycol or sorbitol
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0013
FMN
-
in 40 mM TAPS buffer at pH 8.5 and 25C
0.0317 - 0.5
nitrilotriacetate
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.076
in 30 mM Tris-HCl (pH 8.0), 10 mM FMN, and 2 mM MgCl2, temperature not specified in the publication
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.8
-
pH optimum for NTA consumption in cell extracts. At pH 7.2 and 8.4, 25% of the maximum activity is obtained, and no activity is left at pH 6.0 or 9.0
8.5
-
pH optimum for the purified enzyme
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7 - 9.6
-
35% activity of the purified enzyme remains at pH 7.0 and 9.6
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25
-
12% activity is left at 2C, and no activity is observed at 40C
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.5
calculated from amino acid sequence
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
45000
Chelativorans sp.
-
gel filtration
100000
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
monomer
Chelativorans sp.
-
1 * 45000, SDS-PAGE
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
sitting drop vapor diffusion method, using 1.6 M NH2SO4, 0.1 M NaCl and 0.1 M HEPES pH 7.5
-
sitting drop vapor diffusion method, using 0.2 M magnesium chloride, 0.1 M MES pH 6.0 and 20% PEG 6000
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
enzyme activity is rather unstable in crude cell extract, even in the presence of the protease inhibitors leupeptin and pepstatin
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-70C, crude cell extract, up to 1 month, no loss of activity
-
-70C, purified component B of NTA monooxygenase in 20 mM HEPES buffer with 2 mM dithiothreitol, at pH 7.8, several months, without any significant loss of activity
-
-80C, partially purified, dialyzed enzyme, 3 months, no loss of activity
-
4C, crude cell extract, 48 hours, 50% loss of activity
-
4C, crude extract, 50 mM Tris-HCl, 200 h, 50% loss of activity
-
4C, purified component B of NTA monooxygenase in 20 mM HEPES buffer at pH 7.8, 7 days, 70% loss of activity
-
4C, purified component B of NTA monooxygenase in 20 mM HEPES buffer with 2 mM dithiothreitol and ammonium sulfate to 5% saturation, at pH 7.8, 7 days, 5% loss of activity
-
4C, purified component B of NTA monooxygenase in 20 mM HEPES buffer with 2 mM dithiothreitol, at pH 7.8, 7 days, 20% loss of activity
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ammonium sulfate precipitation, Blue 3GA agarose column chromatography, MonoQ HR 5/5 column chromatography, and hydroxyapatite column chromatography
Chelativorans sp.
-
ammonium sulfate precipitation, phenyl Sepharose column chromatography, TMAE-Fractogel column chromatography, and phenyl Superose column chromatography
-
HisTrap FF column chromatography and Superdex 75 gel filtration
Ni-NTA column chromatography, HiTrap Q anion-exchange column chromatography
-
partially purified by ammonium sulfate precipitation and phenyl agarose column chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli BL21(DE3) Rosetta cells
expressed in Escherichia coli DH5alpha cells
expressed in Escherichia coli JM105 cells
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
induction of nitrilotriacetate-degrading enzyme proceeds faster when the culture is supplied with mixtures of D-glucose and nitrilotriacetate rather than with nitrilotriacetate alone
-
induction of nitrilotriacetate-degrading enzyme proceeds faster when the culture is supplied with mixtures of D-glucose and nitrilotriacetate rather than with nitrilotriacetate alone; synthesis of nitrilotriacetate monooxygenase becomes induced when nitrilotriacetate contributes to more than approximately 1-3% of the total carbon in the substrate mixture supplied
-
-
starvation of nitrilotriacetate-grown cells leads to a loss of nitrilotriacetate monooxygenase protein. A transition from nitrilotriacetate to D-glucose is accompanied by a rapid loss of nitrilotriacetate monooxygenase
synthesis of nitrilotriacetate monooxygenase becomes induced when nitrilotriacetate contributes to more than approximately 1-3% of the total carbon in the substrate mixture supplied
-
Show AA Sequence (234 entries)
Please use the Sequence Search for a certain query.