BRENDA - Enzyme Database
show all sequences of 1.14.11.55

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)
Crystallization (Commentary)
Organism
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 Variants
Commentary
Organism
E140A
residue involved in dimerization, activity similar to wild-type
Sphingopyxis alaskensis
Q127A
residue involved in ectoine binding, about 1% of wild-type activity
Sphingopyxis alaskensis
R139A
residue involved in dimerization, activity similar to wild-type
Sphingopyxis alaskensis
R139A/E140A
residues involved in dimerization, activity similar to wild-type
Sphingopyxis alaskensis
R280A
residue involved in ectoine binding, about 8% of wild-type activity
Sphingopyxis alaskensis
T149A
residue involved in ectoine binding, about 4% of wild-type activity
Sphingopyxis alaskensis
W150A
residue involved in ectoine binding, about 2% of wild-type activity
Sphingopyxis alaskensis
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.7
-
2-oxoglutarate
pH 8.0, 40C
Sphingopyxis alaskensis
9.8
-
ectoine
pH 8.0, 40C
Sphingopyxis alaskensis
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
35290
-
-
Sphingopyxis alaskensis
70730
-
light scattering, recombinant Strep-tagged protein
Sphingopyxis alaskensis
Organism
Organism
UniProt
Commentary
Textmining
Sphingopyxis alaskensis
Q1GNW5
-
-
Sphingopyxis alaskensis DSM 13593
Q1GNW5
-
-
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
ectoine + 2-oxoglutarate + O2
-
740729
Sphingopyxis alaskensis
5-hydroxyectoine + succinate + CO2
-
-
-
?
ectoine + 2-oxoglutarate + O2
-
740729
Sphingopyxis alaskensis DSM 13593
5-hydroxyectoine + succinate + CO2
-
-
-
?
Subunits
Subunits
Commentary
Organism
dimer
2 * 35290, calculated, recombinant Strep-tagged protein
Sphingopyxis alaskensis
Synonyms
Synonyms
Commentary
Organism
Sala_2952
-
Sphingopyxis alaskensis
Crystallization (Commentary) (protein specific)
Crystallization
Organism
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)
Protein Variants
Commentary
Organism
E140A
residue involved in dimerization, activity similar to wild-type
Sphingopyxis alaskensis
Q127A
residue involved in ectoine binding, about 1% of wild-type activity
Sphingopyxis alaskensis
R139A
residue involved in dimerization, activity similar to wild-type
Sphingopyxis alaskensis
R139A/E140A
residues involved in dimerization, activity similar to wild-type
Sphingopyxis alaskensis
R280A
residue involved in ectoine binding, about 8% of wild-type activity
Sphingopyxis alaskensis
T149A
residue involved in ectoine binding, about 4% of wild-type activity
Sphingopyxis alaskensis
W150A
residue involved in ectoine binding, about 2% of wild-type activity
Sphingopyxis alaskensis
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
2.7
-
2-oxoglutarate
pH 8.0, 40C
Sphingopyxis alaskensis
9.8
-
ectoine
pH 8.0, 40C
Sphingopyxis alaskensis
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
35290
-
-
Sphingopyxis alaskensis
70730
-
light scattering, recombinant Strep-tagged protein
Sphingopyxis alaskensis
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
ectoine + 2-oxoglutarate + O2
-
740729
Sphingopyxis alaskensis
5-hydroxyectoine + succinate + CO2
-
-
-
?
ectoine + 2-oxoglutarate + O2
-
740729
Sphingopyxis alaskensis DSM 13593
5-hydroxyectoine + succinate + CO2
-
-
-
?
Subunits (protein specific)
Subunits
Commentary
Organism
dimer
2 * 35290, calculated, recombinant Strep-tagged protein
Sphingopyxis alaskensis
Other publictions for EC
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Synonyms
Temperature Optimum [C]
Temperature Range [C]
Temperature Stability [C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [C] (protein specific)
Temperature Range [C] (protein specific)
Temperature Stability [C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
741719
Tao
Ectoine and 5-hydroxyectoine ...
Virgibacillus halodenitrificans, Virgibacillus halodenitrificans PDB-F2
Appl. Microbiol. Biotechnol.
100
6779-6789
2016
-
-
-
-
-
-
-
-
-
-
-
2
-
2
-
-
-
-
-
-
-
-
2
-
1
-
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-
-
2
-
-
-
-
-
-
-
-
2
-
-
-
-
-
-
-
-
-
1
-
-
1
-
-
742383
Widderich
Strangers in the archaeal wor ...
Nitrosopumilus maritimus
Environ. Microbiol.
18
1227-1248
2016
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-
1
-
-
-
-
-
-
-
1
1
-
7
-
-
1
-
-
-
-
-
1
1
1
-
-
-
-
-
-
-
-
-
-
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-
-
1
-
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-
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-
-
-
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-
1
1
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-
1
-
-
-
-
1
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
741466
Yang
Crystallization and X-ray diff ...
Bacillus pseudofirmus, Bacillus pseudofirmus OF4
Wei Sheng Wu Xue Bao
55
1468-1474
2015
-
-
1
1
-
-
-
-
-
-
-
-
-
5
-
-
-
-
-
-
-
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-
1
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1
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1
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-
-
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-
-
-
-
-
-
-
-
-
-
-
-
-
-
739813
Hoeppner
Overexpression, crystallizatio ...
Sphingopyxis alaskensis
Acta Crystallogr. Sect. F
70
493-496
2014
-
-
1
1
-
-
-
-
-
1
-
-
-
2
-
-
-
-
-
-
-
-
-
-
-
-
-
-
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-
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-
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1
-
1
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1
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-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
740729
Hoeppner
Crystal structure of the ectoi ...
Sphingopyxis alaskensis, Sphingopyxis alaskensis DSM 13593
J. Biol. Chem.
289
29570-29583
2014
-
-
-
1
7
-
-
2
-
-
2
-
-
5
-
-
-
-
-
-
-
-
2
1
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
1
7
-
-
-
-
2
-
-
2
-
-
-
-
-
-
-
-
-
2
1
-
-
-
-
-
-
-
-
-
-
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-
-
-
740868
Widderich
Molecular dynamics simulations ...
Virgibacillus salexigens
J. Mol. Biol.
426
586-600
2014
-
-
-
1
36
-
-
6
-
-
-
-
-
3
-
-
-
-
-
-
-
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1
-
-
-
-
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-
1
36
-
-
-
-
6
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
741304
Widderich
Biochemical properties of ecto ...
Acidiphilium cryptum, Alkalilimnicola ehrlichii, Halomonas elongata, Halomonas elongata DSM 2581, Paenibacillus lautus, Pseudomonas stutzeri, Pseudomonas stutzeri A1501, Sphingopyxis alaskensis, Sphingopyxis alaskensis DSM 13593, Virgibacillus salexigens
PLoS ONE
9
e93809
2014
-
-
-
1
-
-
7
14
-
21
7
20
-
17
-
-
-
-
-
-
-
-
20
7
8
7
7
-
7
7
7
-
-
-
7
-
-
-
-
-
1
-
-
-
7
-
14
-
21
7
20
-
-
-
-
-
-
-
-
20
7
7
7
-
7
7
7
-
7
-
-
-
-
7
7
743087
Widderich
Molecular dynamics simulation ...
Virgibacillus salexigens
J. Mol. Biol.
426
586-600
2014
-
-
1
-
36
-
-
14
-
1
-
1
-
3
-
-
-
-
-
-
-
-
1
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
1
-
-
36
-
-
-
-
14
-
1
-
1
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
740784
Eilert
Synthesis and release of the b ...
Halomonas elongata, Halomonas elongata DSM 2581
J. Biotechnol.
167
85-93
2013
-
1
-
-
-
-
-
-
-
-
-
-
-
3
-
-
-
-
-
-
-
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1
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1
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-
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-
-
-
-
-
-
741265
Reuter
Synthesis of 5-hydroxyectoine ...
Virgibacillus salexigens
PLoS ONE
5
e10647
2010
-
-
1
1
-
-
-
-
-
-
-
-
-
5
-
-
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-
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-
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1
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1
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-
-
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-
739899
Bursy
Synthesis and uptake of the co ...
Streptomyces coelicolor, Streptomyces coelicolor A3(2)
Appl. Environ. Microbiol.
74
7286-7296
2008
-
-
-
-
-
-
1
2
-
1
1
-
-
43
-
-
1
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1
-
4
1
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1
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1
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1
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2
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1
1
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1
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1
-
4
1
1
-
-
-
1
-
-
-
-
1
1
-
-
-
740684
Bursy
Osmotically induced synthesis ...
Virgibacillus salexigens
J. Biol. Chem.
282
31147-31155
2007
-
-
-
-
-
-
1
2
-
1
3
-
-
6
-
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1
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1
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2
1
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1
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1
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1
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2
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1
3
-
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1
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1
-
2
1
1
-
-
-
1
-
-
-
1
-
-
1
-
-
739896
Prabhu
Functional expression of the e ...
Streptomyces anulatus
Appl. Environ. Microbiol.
70
3130-3132
2004
-
1
-
-
-
-
-
-
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3
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1
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1
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1
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1
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