BRENDA - Enzyme Database
show all sequences of 6.3.2.9

Large-scale preparation, purification, and crystallization of UDP-N-acetylmuramoyl-L-alanine: D-glutamate ligase from Escherichia coli

Auger, G.; Martin, L.; Bertrand, J.; Ferrari, P.; Fanchon, E.; Vaganay, S.; Petillot, Y.; van Heijenoort, J.; Blanot, D.; Dideberg, O.; Protein Expr. Purif. 13, 23-29 (1998)

Data extracted from this reference:

Cloned(Commentary)
Commentary
Organism
expressed in Escherichia coli JM83(pMLD58)
Escherichia coli
Crystallization (Commentary)
Crystallization
Organism
hanging-drop vapor diffusion technique
Escherichia coli
Metals/Ions
Metals/Ions
Commentary
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]
Commentary
Organism
46840
-
mass spectrometry
Escherichia coli
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Escherichia coli
-
-
-
Escherichia coli JM83(pMLD58)
-
-
-
Purification (Commentary)
Commentary
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]
Commentary
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
Commentary
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
Commentary
Organism
8.8
-
-
Escherichia coli
Cloned(Commentary) (protein specific)
Commentary
Organism
expressed in Escherichia coli JM83(pMLD58)
Escherichia coli
Crystallization (Commentary) (protein specific)
Crystallization
Organism
hanging-drop vapor diffusion technique
Escherichia coli
Metals/Ions (protein specific)
Metals/Ions
Commentary
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] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
46840
-
mass spectrometry
Escherichia coli
Purification (Commentary) (protein specific)
Commentary
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] (protein specific)
Specific Activity Minimum [µmol/min/mg]
Specific Activity Maximum [µmol/min/mg]
Commentary
Organism
1.772
-
37°C
Escherichia coli
Storage Stability (protein specific)
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] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
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 (protein specific)
pH Optimum Minimum
pH Optimum Maximum
Commentary
Organism
8.8
-
-
Escherichia coli
Other publictions for EC 6.3.2.9
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
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)
746203
Sink
Crystallographic study of pept ...
Escherichia coli
PLoS ONE
11
e0152075
2016
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1
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1
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1
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2
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3
3
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715974
Tomasic
Virtual screening for potentia ...
Escherichia coli
J. Mol. Model.
18
1063-1072
2012
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4
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1
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714902
Sosic
Second-generation sulfonamide ...
Escherichia coli
Eur. J. Med. Chem.
46
2880-2894
2011
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1
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4
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714905
Tomasic
Novel 2-thioxothiazolidin-4-on ...
Escherichia coli
Eur. J. Med. Chem.
46
3964-3975
2011
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3
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715866
Tomasic
Structure-based design of a ne ...
Escherichia coli
J. Med. Chem.
54
4600-4610
2011
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1
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5
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4
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5
5
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714367
Patin
Purification and biochemical c ...
Staphylococcus aureus
Biochimie
92
1793-1800
2010
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1
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3
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1
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715722
Umamaheswari
-
Virtual screening for potentia ...
Leptospira interrogans
J. Chem. Biol.
13
175-187
2010
-
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1
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1
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715847
Zidar
Discovery of novel 5-benzylide ...
Escherichia coli
J. Med. Chem.
53
6584-6594
2010
-
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1
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2
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702636
Turk
Discovery of new inhibitors of ...
Escherichia coli
Bioorg. Med. Chem.
17
1884-1889
2009
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1
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704728
Kristan
New high-throughput fluorimetr ...
Escherichia coli
J. Biomol. Screen.
14
412-418
2009
-
1
1
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2
3
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705031
Simcic
NMR and molecular dynamics stu ...
Escherichia coli
J. Med. Chem.
52
2899-2908
2009
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1
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4
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705227
Perdih
Binding free energy calculatio ...
Escherichia coli
J. Mol. Model.
15
983-996
2009
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3
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690835
Thakur
Ability of PknA, a mycobacteri ...
Mycobacterium tuberculosis, Mycobacterium tuberculosis H37Ra / ATCC 25177
Biochem. J.
415
27-33
2008
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1
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4
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1
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1
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692904
Bratkovic
Peptide inhibitors of MurD and ...
Escherichia coli
J. Basic Microbiol.
48
202-206
2008
-
1
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4
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1
1
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1
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1
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4
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694288
Frlan
Design and synthesis of novel ...
Escherichia coli
Molecules
13
11-30
2008
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25
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6
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6
25
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695027
Perdih
MurD ligase from E. coli: Tetr ...
Escherichia coli
Proteins
15
744-759
2008
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675446
Kotnik
Structural and functional char ...
Escherichia coli
J. Mol. Biol.
370
107-115
2007
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676300
Paradis-Bleau
Selection of peptide inhibitor ...
Pseudomonas aeruginosa
Peptides
27
1693-1700
2006
-
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2
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1
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3
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2
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677175
Humljan
-
A new approach towards peptido ...
Escherichia coli
Tetrahedron
62
10980-10988
2006
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6
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653134
Bouhss
MurC and MurD synthetases of p ...
Escherichia coli, Escherichia coli JM83(pMLD58)
Methods Enzymol.
354
189-196
2002
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