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Literature summary for 6.3.2.9 extracted from

  • Auger, G.; Martin, L.; Bertrand, J.; Ferrari, P.; Fanchon, E.; Vaganay, S.; Petillot, Y.; van Heijenoort, J.; Blanot, D.; Dideberg, O.
    Large-scale preparation, purification, and crystallization of UDP-N-acetylmuramoyl-L-alanine: D-glutamate ligase from Escherichia coli (1998), Protein Expr. Purif., 13, 23-29.
    View publication on PubMed
Search for more literature for 6.3.2.9

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli JM83(pMLD58) Escherichia coli

Crystallization (Commentary)

Crystallization (Comment) Organism
hanging-drop vapor diffusion technique Escherichia coli

Metals/Ions

Metals/Ions Comment Organism Structure
arsenate increases storage stability Escherichia coli
Ca2+ increases storage stability Escherichia coli
Cd2+ abolishes the enzyme activity at 5mM Escherichia coli
Mg2+ increases storage stability Escherichia coli
Mn2+ increases storage stability Escherichia coli
phosphate increases storage stability Escherichia coli
sulfate increases storage stability Escherichia coli
vanadate abolishes the enzyme activity at 5mM Escherichia coli
Zn2+ abolishes the enzyme activity at 5mM Escherichia coli

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
46840
-
 mass spectrometry Escherichia coli

Organism

Organism UniProt Comment Textmining
Escherichia coli
-
-
-
Escherichia coli JM83(pMLD58)
-
-
-

Purification (Commentary)

Purification (Comment) Organism
method that includes DEAE-Trisacryl M and hydroxyapatite-Ultrogel chromatography. Large-scale purification using 2-mercaptoethanol as a reducing agent, results in 200 mg of purified enzyme from 54 l of culture, adducts with one or two molecules of 2-mercaptoethanol are formed. Replacement of 2-mercaptoethanol by dithiothreitol gave 162 mg of enzyme from 36 l of culture yielding no clusters with the reducing agent. Escherichia coli

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
1.772
-
 37°C Escherichia coli

Storage Stability

Storage Stability Organism
-20°C, 20 mM potassium phosphate, pH 7.0, 1 mM 2-mercaptoethanol, 0.1 mM MgCl2, several months, no loss of activity at high enzyme concentrations: 1-10 mg/ml Escherichia coli
-20°C, 20 mM potassium phosphate, pH 7.0, 1 mM dithiothreitol, 0.1 mM MgCl2, several months, no loss of activity at high enzyme concentrations: 1-10 mg/ml Escherichia coli
8°C, 20 mM potassium phosphate, pH 7.0, 1 mM 2-mercaptoethanol, 0.1 mM MgCl2, 1 week, 30% loss of activity Escherichia coli
8°C, 20 mM potassium phosphate, pH 7.0, 1 mM 2-mercaptoethanol, 0.1 mM MgCl2, sodium Hepes, 1 week, 35-43% loss of activity Escherichia coli
8°C, 20 mM potassium phosphate, pH 7.0, 1 mM dithiothreitol, 0.1 mM MgCl2, 1 week, 30% loss of activity Escherichia coli
8°C, 20 mM potassium phosphate, pH 7.0, 1 mM dithiothreitol, 0.1 mM MgCl2, sodium Hepes 1 week, 35-43% loss of activity Escherichia coli
8°C, 20 mM potassium phosphate, pH 8.0, 1 mM 2-mercaptoethanol, 0.1 mM MgCl2, sodium Hepes, 1 week, 35-43% loss of activity Escherichia coli
8°C, 20 mM potassium phosphate, pH 8.0, 1 mM dithiothreitol, 0.1 mM MgCl2, sodium Hepes 1 week, 35-43% loss of activity Escherichia coli

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
additional information
-
 additional information turnover of 6.55 at saturation concentrations of all the substrates: 5 mM ATP, 0.25 mM UDP-N-acetylmuramoyl L-alanine, 0.55 mM D-glutamate Escherichia coli

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8.8
-
-
 Escherichia coli