BRENDA - Enzyme Database
show all sequences of 4.2.1.108

Overproduction, crystallization and X-ray diffraction data analysis of ectoine synthase from the cold-adapted marine bacterium Sphingopyxis alaskensis

Kobus, S.; Widderich, N.; Hoeppner, A.; Bremer, E.; Smits, S.; Acta Crystallogr. Sect. F 71, 1027-1032 (2015)

Data extracted from this reference:

Cloned(Commentary)
Cloned (Commentary)
Organism
gene ectC, recombinant expression of a codon-optimized version of C-terminall Strep II-tagged enzyme in Escherichia coli strain BL21 under control of the tet promoter, which in turn is controlled by the TetR repressor, the genetic expression system can be induced by adding anhydrotetracycline
Sphingopyxis alaskensis
Crystallization (Commentary)
Crystallization (Commentary)
Organism
purified recombinant Strep II-tagged enzyme, sitting drop vapour diffusion method, mixing of 100 nl of 11 mg/ml enzyme solution with 100 nl of reservoir solution containing 0.05 M, calcium acetate, 0.1 M sodium acetate, pH 4.5, and 40% v/v 1,2-propanediol, and equilibration against 0.05 ml of reservoir solution, 12°C, ten weeks, X-ray diffraction structure determination and analysis at 1.0 A resolution, molecular replacement using the crystal structure of Zn2+-containing cupin RemF from Streptomyces resistomycificus (PDB ID 3ht1) as template
Sphingopyxis alaskensis
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
32700
-
recombinant Strep II-tagged enzyme, gel filtration
Sphingopyxis alaskensis
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
(2S)-4-acetamido-2-aminobutanoate
Sphingopyxis alaskensis
-
L-ectoine + H2O
-
-
?
(2S)-4-acetamido-2-aminobutanoate
Sphingopyxis alaskensis DSM 13593 / LMG 18877 / RB2256
-
L-ectoine + H2O
-
-
?
Organism
Organism
UniProt
Commentary
Textmining
Sphingopyxis alaskensis
Q1GNW6
i.e. Sphingomonas alaskensis
-
Sphingopyxis alaskensis DSM 13593 / LMG 18877 / RB2256
Q1GNW6
i.e. Sphingomonas alaskensis
-
Purification (Commentary)
Purification (Commentary)
Organism
recombinant C-terminall Strep II-tagged enzyme from Escherichia coli by affinity chromatography and gel filtration to homogeneity
Sphingopyxis alaskensis
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
(2S)-4-acetamido-2-aminobutanoate
-
746650
Sphingopyxis alaskensis
L-ectoine + H2O
-
-
-
?
(2S)-4-acetamido-2-aminobutanoate
-
746650
Sphingopyxis alaskensis DSM 13593 / LMG 18877 / RB2256
L-ectoine + H2O
-
-
-
?
Subunits
Subunits
Commentary
Organism
dimer
-
Sphingopyxis alaskensis
Synonyms
Synonyms
Commentary
Organism
EctC
-
Sphingopyxis alaskensis
L-ectoine synthase
UniProt
Sphingopyxis alaskensis
SaEctC
-
Sphingopyxis alaskensis
Cloned(Commentary) (protein specific)
Commentary
Organism
gene ectC, recombinant expression of a codon-optimized version of C-terminall Strep II-tagged enzyme in Escherichia coli strain BL21 under control of the tet promoter, which in turn is controlled by the TetR repressor, the genetic expression system can be induced by adding anhydrotetracycline
Sphingopyxis alaskensis
Crystallization (Commentary) (protein specific)
Crystallization
Organism
purified recombinant Strep II-tagged enzyme, sitting drop vapour diffusion method, mixing of 100 nl of 11 mg/ml enzyme solution with 100 nl of reservoir solution containing 0.05 M, calcium acetate, 0.1 M sodium acetate, pH 4.5, and 40% v/v 1,2-propanediol, and equilibration against 0.05 ml of reservoir solution, 12°C, ten weeks, X-ray diffraction structure determination and analysis at 1.0 A resolution, molecular replacement using the crystal structure of Zn2+-containing cupin RemF from Streptomyces resistomycificus (PDB ID 3ht1) as template
Sphingopyxis alaskensis
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
32700
-
recombinant Strep II-tagged enzyme, gel filtration
Sphingopyxis alaskensis
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
(2S)-4-acetamido-2-aminobutanoate
Sphingopyxis alaskensis
-
L-ectoine + H2O
-
-
?
(2S)-4-acetamido-2-aminobutanoate
Sphingopyxis alaskensis DSM 13593 / LMG 18877 / RB2256
-
L-ectoine + H2O
-
-
?
Purification (Commentary) (protein specific)
Commentary
Organism
recombinant C-terminall Strep II-tagged enzyme from Escherichia coli by affinity chromatography and gel filtration to homogeneity
Sphingopyxis alaskensis
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
(2S)-4-acetamido-2-aminobutanoate
-
746650
Sphingopyxis alaskensis
L-ectoine + H2O
-
-
-
?
(2S)-4-acetamido-2-aminobutanoate
-
746650
Sphingopyxis alaskensis DSM 13593 / LMG 18877 / RB2256
L-ectoine + H2O
-
-
-
?
Subunits (protein specific)
Subunits
Commentary
Organism
dimer
-
Sphingopyxis alaskensis
General Information
General Information
Commentary
Organism
evolution
ectoine biosynthetic genes (ectABC) are widely distributed in bacteria, ectoine synthase is a member of the cupin superfamily
Sphingopyxis alaskensis
physiological function
microorganisms that carry the ectoine biosynthetic genes (ectABC) make copious amounts of ectoine as a cell protectant in response to high-osmolarity challenges. Ectoine synthase (EctC) is the key enzyme for the production of this compatible solute and mediates the last step of ectoine biosynthesis. It catalyzes the ring closure of the cyclic ectoine molecule
Sphingopyxis alaskensis
General Information (protein specific)
General Information
Commentary
Organism
evolution
ectoine biosynthetic genes (ectABC) are widely distributed in bacteria, ectoine synthase is a member of the cupin superfamily
Sphingopyxis alaskensis
physiological function
microorganisms that carry the ectoine biosynthetic genes (ectABC) make copious amounts of ectoine as a cell protectant in response to high-osmolarity challenges. Ectoine synthase (EctC) is the key enzyme for the production of this compatible solute and mediates the last step of ectoine biosynthesis. It catalyzes the ring closure of the cyclic ectoine molecule
Sphingopyxis alaskensis
Other publictions for EC 4.2.1.108
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)
749081
Widderich
Biochemistry and crystal stru ...
Sphingopyxis alaskensis, Sphingopyxis alaskensis DSM 13593 / LMG 18877 / RB2256
PLoS ONE
11
e0151285
2016
-
-
1
1
26
-
-
3
-
2
-
2
-
7
-
-
1
-
-
-
-
-
8
2
5
1
-
-
2
1
-
-
-
-
-
-
-
-
1
-
1
26
-
-
-
-
3
-
2
-
2
-
-
-
1
-
-
-
-
8
2
1
-
-
2
1
-
-
-
-
2
2
-
2
2
746650
Kobus
Overproduction, crystallizati ...
Sphingopyxis alaskensis, Sphingopyxis alaskensis DSM 13593 / LMG 18877 / RB2256
Acta Crystallogr. Sect. F
71
1027-1032
2015
-
-
1
1
-
-
-
-
-
-
1
2
-
7
-
-
1
-
-
-
-
-
2
1
4
-
-
-
-
-
-
-
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-
-
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-
1
-
1
-
-
-
-
-
-
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-
1
2
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1
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-
-
-
2
1
-
-
-
-
-
-
-
-
-
2
2
-
-
-
713911
Witt
Unexpected property of ectoine ...
Halomonas elongata, Halomonas elongata DSM 2581
Appl. Microbiol. Biotechnol.
91
113-122
2011
-
-
1
-
1
-
-
1
-
-
1
2
-
5
-
-
1
-
-
-
2
-
11
1
1
1
-
-
-
1
-
-
-
-
-
-
-
-
1
-
-
1
-
-
-
-
1
-
-
1
2
-
-
-
1
-
-
2
-
11
1
1
-
-
-
1
-
-
-
-
3
3
-
-
-
729687
Reshetnikov
Diversity and phylogeny of the ...
Methylarcula marina, Methylarcula marina ML1, Methylobacter marinus, Methylobacter marinus 7C, Methylomicrobium alcaliphilum, Methylomicrobium alcaliphilum ML1, Methylomicrobium kenyense, Methylomicrobium kenyense AMO1, Methylophaga alcalica, Methylophaga alcalica M8, Methylophaga thalassica, Methylophaga thalassica ATCC 33146
Extremophiles
15
653-663
2011
-
-
6
-
-
-
-
-
-
-
-
-
-
19
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
6
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
6
6
-
-
-
690643
Rajan
Characterization and phylogene ...
Bacillus halodurans, Bacillus halodurans TCJAR021
Arch. Microbiol.
190
481-487
2008
-
1
1
-
-
-
-
-
-
-
1
4
-
3
-
-
-
-
-
-
-
-
4
-
1
-
-
-
-
-
-
-
-
-
1
-
-
1
1
-
-
-
-
-
-
-
-
-
-
1
4
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
691437
Rajan
Cloning and heterologous expre ...
Bacillus halodurans, Bacillus halodurans TCJAR021
Biotechnol. Lett.
30
1403-1407
2008
-
1
1
-
-
-
-
-
-
-
1
4
-
3
-
-
-
-
-
-
-
-
4
-
1
-
-
-
-
-
-
-
-
-
1
-
-
1
1
-
-
-
-
-
-
-
-
-
-
1
4
-
-
-
-
-
-
-
-
4
-
-
-
-
-
-
-
-
1
-
-
-
-
-
-
660836
Reshetnikov
Characterization of the ectoin ...
Methylomicrobium alcaliphilum
Arch. Microbiol.
184
286-297
2006
1
-
1
-
-
-
-
-
-
1
-
1
-
3
-
-
-
-
-
1
-
-
2
-
1
1
-
-
-
1
-
-
-
-
-
-
1
-
1
-
-
-
-
-
-
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1
-
1
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-
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-
-
1
-
-
2
-
1
-
-
-
1
-
-
-
-
-
-
-
-
-
660740
Kuhlmann
Osmotically regulated synthesi ...
Bacillus alcalophilus, Chromohalobacter salexigens, no activity in Aneurinibacillus aneurinilyticus, no activity in Aneurinibacillus aneurinilyticus DSM 5562, no activity in Bacillus cereus, no activity in Bacillus cereus DSM 31, no activity in Bacillus circulans, no activity in Bacillus licheniformis, no activity in Bacillus megaterium, no activity in Bacillus subtilis, no activity in Bacillus subtilis JH642, no activity in Bacillus thuringiensis, no activity in Paenibacillus polymyxa, Sporosarcina pasteurii, Sporosarcina psychrophila, Virgibacillus pantothenticus, Virgibacillus salexigens
Appl. Environ. Microbiol.
68
772-783
2002
6
-
2
-
-
-
-
-
-
1
-
12
-
27
-
-
-
-
-
-
-
-
18
-
-
-
-
-
-
-
-
-
-
-
-
-
6
-
2
-
-
-
-
-
-
-
-
-
1
-
12
-
-
-
-
-
-
-
-
18
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
663114
Nakayama
Ectoine, the compatible solute ...
Halomonas elongata, Halomonas elongata OUT30018, no activity in Nicotiana tabacum
Plant Physiol.
122
1239-1247
2000
-
-
1
-
1
-
-
-
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2
-
9
-
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4
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1
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1
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2
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4
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-
-
-
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-
-
-
-
-
-
-
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-
640091
Ono
Characterization of biosynthet ...
Halomonas elongata, Halomonas elongata OUT30018
J. Bacteriol.
181
91-99
1999
-
-
-
-
-
1
-
2
-
2
2
3
-
5
-
-
1
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-
1
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6
1
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3
1
1
-
1
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-
-
1
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-
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-
1
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2
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2
2
3
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1
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-
1
-
6
1
3
1
1
-
1
-
-
1
-
-
-
-
-
-
662773
Louis
Characterization of genes for ...
Marinococcus halophilus
Microbiology
143
1141-1149
1997
1
1
1
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2
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3
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3
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1
1
1
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2
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3
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-
-
-
-
-
-
-
640090
Peters
-
The biosynthesis of ectoine ...
Halomonas elongata, Halorhodospira halochloris
FEMS Microbiol. Lett.
71
157-162
1990
-
-
-
-
-
-
-
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-
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2
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2
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2
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2
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