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Literature summary extracted from

  • Jun, S.Y.; Lewis, K.M.; Youn, B.; Xun, L.; Kang, C.
    Structural and biochemical characterization of EDTA monooxygenase and its physical interaction with a partner flavin reductase (2016), Mol. Microbiol., 100, 989-1003.
    View publication on PubMedView publication on EuropePMC

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
1.14.14.33 to 2.15 A resolution. The structure shows a domain-like insertion into a TIM-barrel, which might serve as a flexible lid for the active site. Docking of MgEDTA2- into EmoA identifies an intricate hydrogen bond network connected to Tyr71, which should lower its pKa. Tyr71, along with nearby Glu70 and a peroxy flavin, facilitates a keto-enol transition of the leaving acetyl group of EDTA. The interaction between EmoA and oxidoreductase EmoB enhances both EmoA and EmoB activities probably through coupled channelling of FMNH2 EDTA-degrading bacterium BNC1

Protein Variants

EC Number Protein Variants Comment Organism
1.14.14.33 E70A complete loss of activtiy EDTA-degrading bacterium BNC1
1.14.14.33 P291A 74.4% of wild-type activity EDTA-degrading bacterium BNC1
1.14.14.33 Q294A 85.6% of wild-type activity EDTA-degrading bacterium BNC1
1.14.14.33 Q58A 51.3% of wild-type activity EDTA-degrading bacterium BNC1
1.14.14.33 R68A 85.3% of wild-type activity EDTA-degrading bacterium BNC1
1.14.14.33 W11A 26.4% of wild-type activity EDTA-degrading bacterium BNC1
1.14.14.33 Y71A complete loss of activtiy EDTA-degrading bacterium BNC1

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.14.14.33 0.0461
-
ethylenediaminetetraacetate wild-type, presence of Mg2+ and oxidoreductase EmoB, pH 7.6, temperature not specified in the publication EDTA-degrading bacterium BNC1
1.14.14.33 0.047
-
ethylenediaminetetraacetate mutant R68A, presence of Mg2+, pH 7.6, temperature not specified in the publication EDTA-degrading bacterium BNC1
1.14.14.33 0.0492
-
ethylenediaminetetraacetate wild-type, presence of Mg2+, pH 7.6, temperature not specified in the publication EDTA-degrading bacterium BNC1
1.14.14.33 0.0615
-
ethylenediaminetetraacetate wild-type, presence of Mg2+ and flavin reductase from Escherichia coli, pH 7.6, temperature not specified in the publication EDTA-degrading bacterium BNC1
1.14.14.33 0.069
-
ethylenediaminetetraacetate mutant Q294A, presence of Mg2+, pH 7.6, temperature not specified in the publication EDTA-degrading bacterium BNC1
1.14.14.33 0.198
-
ethylenediaminetetraacetate mutant W11A, presence of Mg2+, pH 7.6, temperature not specified in the publication EDTA-degrading bacterium BNC1
1.14.14.33 0.49
-
ethylenediaminetetraacetate mutant Q58A, presence of Mg2+, pH 7.6, temperature not specified in the publication EDTA-degrading bacterium BNC1
1.14.14.33 0.521
-
ethylenediaminetetraacetate mutant P291A, presence of Mg2+, pH 7.6, temperature not specified in the publication EDTA-degrading bacterium BNC1

Organism

EC Number Organism UniProt Comment Textmining
1.14.14.33 EDTA-degrading bacterium BNC1 Q9F9T3
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
1.14.14.33 ethylenediaminetetraacetate + 2 FMNH2 + 2 O2
-
EDTA-degrading bacterium BNC1 ethylenediamine-N,N'-diacetate + 2 glyoxylate + 2 FMN + 2 H2O
-
?

Subunits

EC Number Subunits Comment Organism
1.14.14.33 dimer light scattering experiments EDTA-degrading bacterium BNC1

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
1.14.14.33 4.5
-
ethylenediaminetetraacetate mutant W11A, presence of Mg2+, pH 7.6, temperature not specified in the publication EDTA-degrading bacterium BNC1
1.14.14.33 8.8
-
ethylenediaminetetraacetate mutant Q58A, presence of Mg2+, pH 7.6, temperature not specified in the publication EDTA-degrading bacterium BNC1
1.14.14.33 12.8
-
ethylenediaminetetraacetate mutant P291A, presence of Mg2+, pH 7.6, temperature not specified in the publication EDTA-degrading bacterium BNC1
1.14.14.33 13.2
-
ethylenediaminetetraacetate wild-type, presence of Mg2+ and flavin reductase from Escherichia coli, pH 7.6, temperature not specified in the publication EDTA-degrading bacterium BNC1
1.14.14.33 14.7
-
ethylenediaminetetraacetate mutant Q294A, presence of Mg2+, pH 7.6, temperature not specified in the publication EDTA-degrading bacterium BNC1
1.14.14.33 14.7
-
ethylenediaminetetraacetate mutant R68A, presence of Mg2+, pH 7.6, temperature not specified in the publication EDTA-degrading bacterium BNC1
1.14.14.33 17.2
-
ethylenediaminetetraacetate wild-type, presence of Mg2+, pH 7.6, temperature not specified in the publication EDTA-degrading bacterium BNC1
1.14.14.33 47.1
-
ethylenediaminetetraacetate wild-type, presence of Mg2+ and oxidoreductase EmoB, pH 7.6, temperature not specified in the publication EDTA-degrading bacterium BNC1

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
1.14.14.33 20
-
ethylenediaminetetraacetate mutant P291A, presence of Mg2+, pH 7.6, temperature not specified in the publication EDTA-degrading bacterium BNC1
1.14.14.33 20
-
ethylenediaminetetraacetate mutant Q58A, presence of Mg2+, pH 7.6, temperature not specified in the publication EDTA-degrading bacterium BNC1
1.14.14.33 20
-
ethylenediaminetetraacetate mutant W11A, presence of Mg2+, pH 7.6, temperature not specified in the publication EDTA-degrading bacterium BNC1
1.14.14.33 31
-
ethylenediaminetetraacetate mutant R68A, presence of Mg2+, pH 7.6, temperature not specified in the publication EDTA-degrading bacterium BNC1
1.14.14.33 210
-
ethylenediaminetetraacetate mutant Q294A, presence of Mg2+, pH 7.6, temperature not specified in the publication EDTA-degrading bacterium BNC1
1.14.14.33 350
-
ethylenediaminetetraacetate wild-type, presence of Mg2+, pH 7.6, temperature not specified in the publication EDTA-degrading bacterium BNC1