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)
EC Number
Cloned (Commentary)
Organism
4.2.1.108
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)
EC Number
Crystallization (Commentary)
Organism
4.2.1.108
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]
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
4.2.1.108
32700
-
recombinant Strep II-tagged enzyme, gel filtration
Sphingopyxis alaskensis
Natural Substrates/ Products (Substrates)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
4.2.1.108
(2S)-4-acetamido-2-aminobutanoate
Sphingopyxis alaskensis
-
L-ectoine + H2O
-
-
?
4.2.1.108
(2S)-4-acetamido-2-aminobutanoate
Sphingopyxis alaskensis DSM 13593 / LMG 18877 / RB2256
-
L-ectoine + H2O
-
-
?
Organism
EC Number
Organism
UniProt
Commentary
Textmining
4.2.1.108
Sphingopyxis alaskensis
Q1GNW6
i.e. Sphingomonas alaskensis
-
4.2.1.108
Sphingopyxis alaskensis DSM 13593 / LMG 18877 / RB2256
Q1GNW6
i.e. Sphingomonas alaskensis
-
Purification (Commentary)
EC Number
Purification (Commentary)
Organism
4.2.1.108
recombinant C-terminall Strep II-tagged enzyme from Escherichia coli by affinity chromatography and gel filtration to homogeneity
Sphingopyxis alaskensis
Substrates and Products (Substrate)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
Substrate Product ID
4.2.1.108
(2S)-4-acetamido-2-aminobutanoate
-
746650
Sphingopyxis alaskensis
L-ectoine + H2O
-
-
-
?
4.2.1.108
(2S)-4-acetamido-2-aminobutanoate
-
746650
Sphingopyxis alaskensis DSM 13593 / LMG 18877 / RB2256
L-ectoine + H2O
-
-
-
?
Subunits
EC Number
Subunits
Commentary
Organism
4.2.1.108
dimer
-
Sphingopyxis alaskensis
Synonyms
EC Number
Synonyms
Commentary
Organism
4.2.1.108
EctC
-
Sphingopyxis alaskensis
4.2.1.108
L-ectoine synthase
UniProt
Sphingopyxis alaskensis
4.2.1.108
SaEctC
-
Sphingopyxis alaskensis
Cloned(Commentary) (protein specific)
EC Number
Commentary
Organism
4.2.1.108
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)
EC Number
Crystallization
Organism
4.2.1.108
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)
EC Number
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
4.2.1.108
32700
-
recombinant Strep II-tagged enzyme, gel filtration
Sphingopyxis alaskensis
Natural Substrates/ Products (Substrates) (protein specific)
EC Number
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ID
4.2.1.108
(2S)-4-acetamido-2-aminobutanoate
Sphingopyxis alaskensis
-
L-ectoine + H2O
-
-
?
4.2.1.108
(2S)-4-acetamido-2-aminobutanoate
Sphingopyxis alaskensis DSM 13593 / LMG 18877 / RB2256
-
L-ectoine + H2O
-
-
?
Purification (Commentary) (protein specific)
EC Number
Commentary
Organism
4.2.1.108
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)
EC Number
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ID
4.2.1.108
(2S)-4-acetamido-2-aminobutanoate
-
746650
Sphingopyxis alaskensis
L-ectoine + H2O
-
-
-
?
4.2.1.108
(2S)-4-acetamido-2-aminobutanoate
-
746650
Sphingopyxis alaskensis DSM 13593 / LMG 18877 / RB2256
L-ectoine + H2O
-
-
-
?
Subunits (protein specific)
EC Number
Subunits
Commentary
Organism
4.2.1.108
dimer
-
Sphingopyxis alaskensis
General Information
EC Number
General Information
Commentary
Organism
4.2.1.108
evolution
ectoine biosynthetic genes (ectABC) are widely distributed in bacteria, ectoine synthase is a member of the cupin superfamily
Sphingopyxis alaskensis
4.2.1.108
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)
EC Number
General Information
Commentary
Organism
4.2.1.108
evolution
ectoine biosynthetic genes (ectABC) are widely distributed in bacteria, ectoine synthase is a member of the cupin superfamily
Sphingopyxis alaskensis
4.2.1.108
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