Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary extracted from

  • Kaplun, A.; Binshtein, E.; Vyazmensky, M.; Steinmatz, A.; Barak, Z.; Chipman, D.M.; Tittmann, K.; Shaanan, B.
    Glyoxylate carboligase lacks the canonical active site glutamate of thiamine-dependent enzymes (2008), Nat. Chem. Biol., 4, 113-118.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.1.1.47 expression in Escherichia coli Escherichia coli K-12

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
4.1.1.47 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

EC Number Protein Variants Comment Organism
4.1.1.47 E52Q site-directed mutagenesis Escherichia coli K-12
4.1.1.47 V51D site-directed mutagenesis Escherichia coli K-12
4.1.1.47 V51E site-directed mutagenesis Escherichia coli K-12
4.1.1.47 V51S site-directed mutagenesis Escherichia coli K-12

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
4.1.1.47 0.2
-
 glyoxylate mutant V51D Escherichia coli K-12
4.1.1.47 0.54
-
 glyoxylate mutant V51E Escherichia coli K-12
4.1.1.47 0.9
-
 glyoxylate wild-type enzyme Escherichia coli K-12
4.1.1.47 1.1
-
 glyoxylate mutant V51S Escherichia coli K-12
4.1.1.47 1.2
-
 glyoxylate mutant E52Q Escherichia coli K-12

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
4.1.1.47 Mg2+
-
 Escherichia coli K-12

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
4.1.1.47 2 glyoxylate Escherichia coli K-12
-
 tartronate semialdehyde + CO2
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
4.1.1.47 Escherichia coli K-12 P0AEP7
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.1.1.47 nickel affinity chromatography Escherichia coli K-12

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
4.1.1.47 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
4.1.1.47 2.7
-
 mutant V51E is about seven times slower than the wild-type enzyme Escherichia coli K-12
4.1.1.47 17.1
-
 mutant V51S is nearly as active as the wild-type enzyme Escherichia coli K-12
4.1.1.47 17.5
-
 wild-type enzyme Escherichia coli K-12
4.1.1.47 18.3
-
 mutant E52Q Escherichia coli K-12

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.1.1.47 2 glyoxylate
-
 Escherichia coli K-12 tartronate semialdehyde + CO2
-
 ?

Subunits

EC Number Subunits Comment Organism
4.1.1.47 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

EC Number Synonyms Comment Organism
4.1.1.47 GCL
-
 Escherichia coli K-12
4.1.1.47 Glyoxylate carboligase
-
 Escherichia coli K-12

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
4.1.1.47 0.25
-
 glyoxylate mutant V51D Escherichia coli K-12
4.1.1.47 2.9
-
 glyoxylate mutant V51E Escherichia coli K-12
4.1.1.47 18.5
-
 glyoxylate mutant V51S Escherichia coli K-12
4.1.1.47 18.9
-
 glyoxylate wild-type enzyme Escherichia coli K-12
4.1.1.47 19.7
-
 glyoxylate mutant E52Q Escherichia coli K-12

Cofactor

EC Number Cofactor Comment Organism Structure
4.1.1.47 FAD
-
 Escherichia coli K-12
4.1.1.47 thiamine diphosphate
-
 Escherichia coli K-12

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
4.1.1.47 1.3
-
 glyoxylate mutant V51D Escherichia coli K-12
4.1.1.47 5.3
-
 glyoxylate mutant V51E Escherichia coli K-12
4.1.1.47 16.5
-
 glyoxylate mutant E52Q Escherichia coli K-12
4.1.1.47 17.3
-
 glyoxylate mutant V51S Escherichia coli K-12
4.1.1.47 21
-
 glyoxylate wild-type enzyme Escherichia coli K-12