Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli | Escherichia coli K-12 |
Crystallization (Comment) | Organism |
---|---|
glyoxylate carboligase crystallizes with 6 monomers (a tetramer and a dimer) in an asymmetric unit, vapor diffusion hanging-drop, 2-4 microlitre of protein solution (5-15 mg/ml, 100 micromolar ThDP, 10 micromolar FAD, 1mM MgCl2 and 10 mM quinone Q0) are mixed with equal volume of reservoir solution (0.5% PEG6000, 0.5M NaCl, 40 mM DTT), pH 8.00, temperature 294K, space group P41212, resolution 2.70 A | Escherichia coli K-12 |
Protein Variants | Comment | Organism |
---|---|---|
E52Q | site-directed mutagenesis | Escherichia coli K-12 |
V51D | site-directed mutagenesis | Escherichia coli K-12 |
V51E | site-directed mutagenesis | Escherichia coli K-12 |
V51S | site-directed mutagenesis | Escherichia coli K-12 |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.2 | - |
glyoxylate | mutant V51D | Escherichia coli K-12 | |
0.54 | - |
glyoxylate | mutant V51E | Escherichia coli K-12 | |
0.9 | - |
glyoxylate | wild-type enzyme | Escherichia coli K-12 | |
1.1 | - |
glyoxylate | mutant V51S | Escherichia coli K-12 | |
1.2 | - |
glyoxylate | mutant E52Q | Escherichia coli K-12 |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | - |
Escherichia coli K-12 |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
2 glyoxylate | Escherichia coli K-12 | - |
tartronate semialdehyde + CO2 | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli K-12 | P0AEP7 | - |
- |
Purification (Comment) | Organism |
---|---|
nickel affinity chromatography | Escherichia coli K-12 |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
0.2 | - |
mutant V51D catalyzes the formation of the product nearly two orders of magnitude more slowly than the wild-type enzyme | Escherichia coli K-12 |
2.7 | - |
mutant V51E is about seven times slower than the wild-type enzyme | Escherichia coli K-12 |
17.1 | - |
mutant V51S is nearly as active as the wild-type enzyme | Escherichia coli K-12 |
17.5 | - |
wild-type enzyme | Escherichia coli K-12 |
18.3 | - |
mutant E52Q | Escherichia coli K-12 |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2 glyoxylate | - |
Escherichia coli K-12 | tartronate semialdehyde + CO2 | - |
? |
Subunits | Comment | Organism |
---|---|---|
homotetramer | the apparent functional unit is a dimer with a pair of identical thiamine diphosphate binding sites at the dimer interface | Escherichia coli K-12 |
Synonyms | Comment | Organism |
---|---|---|
GCL | - |
Escherichia coli K-12 |
Glyoxylate carboligase | - |
Escherichia coli K-12 |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.25 | - |
glyoxylate | mutant V51D | Escherichia coli K-12 | |
2.9 | - |
glyoxylate | mutant V51E | Escherichia coli K-12 | |
18.5 | - |
glyoxylate | mutant V51S | Escherichia coli K-12 | |
18.9 | - |
glyoxylate | wild-type enzyme | Escherichia coli K-12 | |
19.7 | - |
glyoxylate | mutant E52Q | Escherichia coli K-12 |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
FAD | - |
Escherichia coli K-12 | |
thiamine diphosphate | - |
Escherichia coli K-12 |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.3 | - |
glyoxylate | mutant V51D | Escherichia coli K-12 | |
5.3 | - |
glyoxylate | mutant V51E | Escherichia coli K-12 | |
16.5 | - |
glyoxylate | mutant E52Q | Escherichia coli K-12 | |
17.3 | - |
glyoxylate | mutant V51S | Escherichia coli K-12 | |
21 | - |
glyoxylate | wild-type enzyme | Escherichia coli K-12 |