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Literature summary for 5.4.99.1 extracted from

  • Chen, H.P.; Marsh, E.N.G.
    Adenosylcobalamin-dependent glutamate mutase: examination of substrate and coenzyme binding in an engineered fusion protein possessing simplified subunit structure and kinetic properties (1997), Biochemistry, 36, 14939-14945.
    View publication on PubMed

Protein Variants

Protein Variants Comment Organism
additional information the S subunit is genetically fused to the C-terminus of the E subunit through an 11 amino acid 5-Gly linker segment. The affinity of adenosylcobalamine is unchanged, but the turnover-number and the Km-value for Glu in the conversion of L-Glu to (2S,3S)-3-methylaspartate are decreased by about a third Clostridium cochlearium

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.14
-
L-threo-3-methylaspartate genetically engineered enzyme with S subunit fused to the C-terminus of the E-subunit through an 11 amino acid (Gly-Gln)5-Gly linker segment Clostridium cochlearium
0.58
-
L-Glu genetically engineered enzyme with S subunit fused to the C-terminus of the E-subunit through an 11 amino acid (Gly-Gln)5-Gly linker segment Clostridium cochlearium

Organism

Organism UniProt Comment Textmining
Clostridium cochlearium
-
recombinant enzyme expressed in Escherichia coli
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-Glu
-
Clostridium cochlearium threo-3-Methylaspartate (2S,3S)-3-methylaspartate ?
L-threo-3-methylaspartate
-
Clostridium cochlearium L-glutamate
-
?

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
5.8
-
L-Glu genetically engineered enzyme with S subunit fused to the C-terminus of the E subunit through an 11 amino acid (Gly-Gln)5-Gly linker segment Clostridium cochlearium
5.8
-
(2S,3S)-3-methylaspartate genetically engineered enzyme with S subunit fused to the C-terminus of the E subunit through an 11 amino acid (Gly-Gln)5-Gly linker segment Clostridium cochlearium

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
protein with the S subunit genetically fused to the C-terminus of the E-subunit through an 11 amino acid (Gly-Gln)5-Gly linker segment Clostridium cochlearium
8 8.5 wild type enzyme Clostridium cochlearium

Cofactor

Cofactor Comment Organism Structure
Cobalamin the weakly associated subunits E and S combine to form the coenzyme binding site. Interactions between the protein and the adenosyl moiety do not serve to weaken the cobalt-carbon bond in the ground state, variation of the apparent Km-value for adenosylcobalamine with protein concentration. Km: 0.0055 mM in the reaction with L-Glu, Km: 0.0031 mM in the reaction with (2S,3S)-3-methylaspartate, genetically engineered enzyme with S subunit fused to the C-terminus of the E subunit through an 11 amino acid (Gly-Gln)5-Gly linker segment Clostridium cochlearium