BRENDA - Enzyme Database

Crystal structure of the ectoine hydroxylase, a snapshot of the active site

Hoeppner, A.; Widderich, N.; Lenders, M.; Bremer, E.; Smits, S.H.; J. Biol. Chem. 289, 29570-29583 (2014)

Data extracted from this reference:

Crystallization (Commentary)
EC Number
Crystallization (Commentary)
Organism
1.14.11.55
crystal structure in its apo-form, in complex with iron, and in complex with iron, cosubstrate 2-oxoglutarate, and 5-hydroxyectoine. The iron and 2-oxoglutarate ligands are bound within the active site in a fashion similar to that found in other members of the dioxygenase superfamily. 5-Hydroxyectoine is bound by residues residues His144, His245, and Asp146 forming the 2-His-1-carboxylate facial triad
Sphingopyxis alaskensis
Engineering
EC Number
Protein Variants
Commentary
Organism
1.14.11.55
E140A
residue involved in dimerization, activity similar to wild-type
Sphingopyxis alaskensis
1.14.11.55
Q127A
residue involved in ectoine binding, about 1% of wild-type activity
Sphingopyxis alaskensis
1.14.11.55
R139A
residue involved in dimerization, activity similar to wild-type
Sphingopyxis alaskensis
1.14.11.55
R139A/E140A
residues involved in dimerization, activity similar to wild-type
Sphingopyxis alaskensis
1.14.11.55
R280A
residue involved in ectoine binding, about 8% of wild-type activity
Sphingopyxis alaskensis
1.14.11.55
T149A
residue involved in ectoine binding, about 4% of wild-type activity
Sphingopyxis alaskensis
1.14.11.55
W150A
residue involved in ectoine binding, about 2% of wild-type activity
Sphingopyxis alaskensis
KM Value [mM]
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.14.11.55
2.7
-
2-oxoglutarate
pH 8.0, 40C
Sphingopyxis alaskensis
1.14.11.55
9.8
-
ectoine
pH 8.0, 40C
Sphingopyxis alaskensis
Molecular Weight [Da]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.14.11.55
35290
-
-
Sphingopyxis alaskensis
1.14.11.55
70730
-
light scattering, recombinant Strep-tagged protein
Sphingopyxis alaskensis
Organism
EC Number
Organism
UniProt
Commentary
Textmining
1.14.11.55
Sphingopyxis alaskensis
Q1GNW5
-
-
1.14.11.55
Sphingopyxis alaskensis DSM 13593
Q1GNW5
-
-
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
1.14.11.55
ectoine + 2-oxoglutarate + O2
-
740729
Sphingopyxis alaskensis
5-hydroxyectoine + succinate + CO2
-
-
-
?
1.14.11.55
ectoine + 2-oxoglutarate + O2
-
740729
Sphingopyxis alaskensis DSM 13593
5-hydroxyectoine + succinate + CO2
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
1.14.11.55
dimer
2 * 35290, calculated, recombinant Strep-tagged protein
Sphingopyxis alaskensis
Synonyms
EC Number
Synonyms
Commentary
Organism
1.14.11.55
Sala_2952
-
Sphingopyxis alaskensis
Crystallization (Commentary) (protein specific)
EC Number
Crystallization
Organism
1.14.11.55
crystal structure in its apo-form, in complex with iron, and in complex with iron, cosubstrate 2-oxoglutarate, and 5-hydroxyectoine. The iron and 2-oxoglutarate ligands are bound within the active site in a fashion similar to that found in other members of the dioxygenase superfamily. 5-Hydroxyectoine is bound by residues residues His144, His245, and Asp146 forming the 2-His-1-carboxylate facial triad
Sphingopyxis alaskensis
Engineering (protein specific)
EC Number
Protein Variants
Commentary
Organism
1.14.11.55
E140A
residue involved in dimerization, activity similar to wild-type
Sphingopyxis alaskensis
1.14.11.55
Q127A
residue involved in ectoine binding, about 1% of wild-type activity
Sphingopyxis alaskensis
1.14.11.55
R139A
residue involved in dimerization, activity similar to wild-type
Sphingopyxis alaskensis
1.14.11.55
R139A/E140A
residues involved in dimerization, activity similar to wild-type
Sphingopyxis alaskensis
1.14.11.55
R280A
residue involved in ectoine binding, about 8% of wild-type activity
Sphingopyxis alaskensis
1.14.11.55
T149A
residue involved in ectoine binding, about 4% of wild-type activity
Sphingopyxis alaskensis
1.14.11.55
W150A
residue involved in ectoine binding, about 2% of wild-type activity
Sphingopyxis alaskensis
KM Value [mM] (protein specific)
EC Number
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
1.14.11.55
2.7
-
2-oxoglutarate
pH 8.0, 40C
Sphingopyxis alaskensis
1.14.11.55
9.8
-
ectoine
pH 8.0, 40C
Sphingopyxis alaskensis
Molecular Weight [Da] (protein specific)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
1.14.11.55
35290
-
-
Sphingopyxis alaskensis
1.14.11.55
70730
-
light scattering, recombinant Strep-tagged protein
Sphingopyxis alaskensis
Substrates and Products (Substrate) (protein specific)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
1.14.11.55
ectoine + 2-oxoglutarate + O2
-
740729
Sphingopyxis alaskensis
5-hydroxyectoine + succinate + CO2
-
-
-
?
1.14.11.55
ectoine + 2-oxoglutarate + O2
-
740729
Sphingopyxis alaskensis DSM 13593
5-hydroxyectoine + succinate + CO2
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
1.14.11.55
dimer
2 * 35290, calculated, recombinant Strep-tagged protein
Sphingopyxis alaskensis