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Literature summary extracted from

  • Uetz, T.; Schneider, R.; Snozzi, M.; Egli, T.
    Purification and characterization of a two-component monooxygenase that hydroxylates nitrilotriacetate from "Chelatobacter" strain ATCC 29600 (1992), J. Bacteriol., 174, 1179-1188.
    View publication on PubMedView publication on EuropePMC

Activating Compound

EC Number Activating Compound Comment Organism Structure
1.14.14.10 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 Aminobacter aminovorans
1.14.14.10 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 Aminobacter aminovorans
1.14.14.10 additional information the activity is increased neither by additions of 20% (w/v) polyethylene glycol or sorbitol Aminobacter aminovorans

Inhibitors

EC Number Inhibitors Comment Organism Structure
1.14.14.10 5,5'-dithiobis(2-nitrobenzoate) 67% residual activity at 1 mM Aminobacter aminovorans
1.14.14.10 Ca2+ 5% residual activity at 10 mM Aminobacter aminovorans
1.14.14.10 EDTA 3% residual activity at 10 mM Aminobacter aminovorans
1.14.14.10 FAD 24% residual activity at 0.1 mM Aminobacter aminovorans
1.14.14.10 NaCl 54% residual activity at 250 mM Aminobacter aminovorans
1.14.14.10 p-hydroxymercuribenzoate 12% residual activity at 1 mM Aminobacter aminovorans

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.5.1.42 0.0013
-
FMN pH 7.8, 25°C Aminobacter aminovorans
1.5.1.42 0.25
-
NADH pH 7.8, 25°C Aminobacter aminovorans
1.14.14.10 0.0013
-
FMN in 40 mM TAPS buffer at pH 8.5 and 25°C Aminobacter aminovorans
1.14.14.10 0.5
-
nitrilotriacetate in 40 mM TAPS buffer at pH 8.5 and 25°C Aminobacter aminovorans

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
1.14.14.10 Co2+ may replace Mg2+, enzyme activity is reduced to 80% of that with MgCl2 Aminobacter aminovorans
1.14.14.10 Mg2+ required, 2 mM used in assay conditions Aminobacter aminovorans
1.14.14.10 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 Aminobacter aminovorans
1.14.14.10 additional information no nitrilotriacetate consumption is observed with Ca2+,Fe2+, Fe3+, Zn2+, Cu2+, or Ni2+ Aminobacter aminovorans
1.14.14.10 additional information the enzyme contains less than 0.15 atom of Fe per mol of protein, indicating that neither Fe-sulfur clusters nor cytochromes are present Aminobacter aminovorans

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
1.14.14.10 36000
-
2 * 36000, component B of NTA monooxygenase, SDS-PAGE Aminobacter aminovorans

Organism

EC Number Organism UniProt Comment Textmining
1.5.1.42 Aminobacter aminovorans
-
i.e. Chelatobacter heintzii
-
1.14.14.10 Aminobacter aminovorans
-
-
-
1.14.14.10 Aminobacter aminovorans ATCC 29600
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
1.5.1.42
-
Aminobacter aminovorans
1.14.14.10 ammonium sulfate precipitation, phenyl Sepharose column chromatography, TMAE-Fractogel column chromatography, and phenyl Superose column chromatography Aminobacter aminovorans

Storage Stability

EC Number Storage Stability Organism
1.5.1.42 -70°C, in presence of 2 mM dithiothreitol, stable over several months Aminobacter aminovorans
1.14.14.10 -70°C, 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 Aminobacter aminovorans
1.14.14.10 4°C, crude extract, 50 mM Tris-HCl, 200 h, 50% loss of activity Aminobacter aminovorans
1.14.14.10 4°C, purified component B of NTA monooxygenase in 20 mM HEPES buffer at pH 7.8, 7 days, 70% loss of activity Aminobacter aminovorans
1.14.14.10 4°C, 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 Aminobacter aminovorans
1.14.14.10 4°C, 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 Aminobacter aminovorans

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.5.1.42 FMN + NADH + H+
-
Aminobacter aminovorans FMNH2 + NAD+
-
?
1.5.1.42 FMN + NADH + H+
-
Aminobacter aminovorans ATCC 29600 FMNH2 + NAD+
-
?
1.14.14.10 additional information less than 2% reactivity with N-methyl-iminodiacetate, N-acetamido-iminodiacetate, N-hydroxyethyl-iminodiacetate, EDTA, trimethylamine, triethanolamine, dimethylamine, dimethylglycine, methylamine, sarcosine, anthranilate, betaine, N-methylglutamate, N-methylalanine, N-methylaspartate, N-acetylglutamate, N-methylphenylalanine, N-methylleucine, N-methyltryptophane, glutamate, aspartate, citrate, proline, imidazoleacetate, asparagine, and ethylenediamine-N,N'-diacetate Aminobacter aminovorans ?
-
?
1.14.14.10 additional information less than 2% reactivity with N-methyl-iminodiacetate, N-acetamido-iminodiacetate, N-hydroxyethyl-iminodiacetate, EDTA, trimethylamine, triethanolamine, dimethylamine, dimethylglycine, methylamine, sarcosine, anthranilate, betaine, N-methylglutamate, N-methylalanine, N-methylaspartate, N-acetylglutamate, N-methylphenylalanine, N-methylleucine, N-methyltryptophane, glutamate, aspartate, citrate, proline, imidazoleacetate, asparagine, and ethylenediamine-N,N'-diacetate Aminobacter aminovorans ATCC 29600 ?
-
?
1.14.14.10 nitrilotriacetate + FMNH2 + H+ + O2 specific substrate Aminobacter aminovorans iminodiacetate + glyoxylate + FMN + H2O
-
?
1.14.14.10 nitrilotriacetate + FMNH2 + H+ + O2 specific substrate Aminobacter aminovorans ATCC 29600 iminodiacetate + glyoxylate + FMN + H2O
-
?

Subunits

EC Number Subunits Comment Organism
1.14.14.10 homodimer 2 * 36000, component B of NTA monooxygenase, SDS-PAGE Aminobacter aminovorans

Synonyms

EC Number Synonyms Comment Organism
1.14.14.10 component B of NTA monooxygenase
-
Aminobacter aminovorans
1.14.14.10 NTA monooxygenase
-
Aminobacter aminovorans
1.14.14.10 NTA-MO
-
Aminobacter aminovorans

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
1.5.1.42 25
-
assay at Aminobacter aminovorans
1.14.14.10 25
-
12% activity is left at 2°C, and no activity is observed at 40°C Aminobacter aminovorans

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
1.5.1.42 7.8
-
assay at Aminobacter aminovorans
1.14.14.10 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 Aminobacter aminovorans
1.14.14.10 8.5
-
pH optimum for the purified enzyme Aminobacter aminovorans

pH Range

EC Number pH Minimum pH Maximum Comment Organism
1.14.14.10 7 9.6 35% activity of the purified enzyme remains at pH 7.0 and 9.6 Aminobacter aminovorans

Cofactor

EC Number Cofactor Comment Organism Structure
1.14.14.10 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 Aminobacter aminovorans
1.14.14.10 NADH NADPH cannot replace NADH Aminobacter aminovorans