Literature summary for 4.2.99.20 extracted from

Sun, Y.; Yin, S.; Feng, Y.; Li, J.; Zhou, J.; Liu, C.; Zhu, G.; Guo, Z.
Molecular basis of the general base catalysis of an alpha/beta-hydrolase catalytic triad (2014), J. Biol. Chem., 289, 15867-15879.

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Cloned(Commentary)

Cloned (Comment)	Organism
-	Escherichia coli

Crystallization (Commentary)

Crystallization (Comment)	Organism
to 1.45 A resolution. The nucleophilicity of the catalytic serine-histidine-aspartate triad is shielded and its catalytic role is limited to being a specific general base by an open-closed conformational change	Escherichia coli

Protein Variants

Protein Variants	Comment	Organism
F153A	residue involved in open-closed transition, mutation leads to large decrease in enzymatic activity	Escherichia coli
V152A	residue involved in open-closed transition, mutation leads to large decrease in enzymatic activity	Escherichia coli
V152G	residue involved in open-closed transition, mutation leads to large decrease in enzymatic activity	Escherichia coli
V152G/F153A	inactive	Escherichia coli
W147A/Y148A	inactive	Escherichia coli
Y148A	residue involved in open-closed transition, mutation leads to large decrease in enzymatic activity	Escherichia coli
Y148F	residue involved in open-closed transition, mutation leads to large decrease in enzymatic activity	Escherichia coli

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.013	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	wild-type, pH 7.0, temperature not specified in the publication	Escherichia coli
0.036	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	mutant Y148F, pH 7.0, temperature not specified in the publication	Escherichia coli
0.05	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	mutant Y148A, pH 7.0, temperature not specified in the publication	Escherichia coli
0.07	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	mutant F153A, pH 7.0, temperature not specified in the publication	Escherichia coli
0.16	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	mutant V152G, pH 7.0, temperature not specified in the publication	Escherichia coli
0.3	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	mutant V152A, pH 7.0, temperature not specified in the publication	Escherichia coli

Organism

Organism	UniProt	Comment	Textmining
Escherichia coli	P37355	-	-

Reaction

Reaction	Comment	Organism	Reaction ID
5-enolpyruvoyl-6-hydroxy-2-succinylcyclohex-3-ene-1-carboxylate = (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate + pyruvate	the nucleophilicity of the catalyitc serine-histidine-aspartate triad is shielded and its catalytic role is limited to being a specific general base by an open-closed conformational change. The enzyme adopts an open conformation without a functional triad in its ligand-free form and a closed conformation with a fully functional catalytic triad in the presence of its reaction product. The open-to-closed conformational transition involves movement of half of the alpha-helical cap domain, which causes extensive structural changes in the apha/beta-domain and forces the side chainof the triad histidine to adopt an energetically disfavored gauche conformation to form the functional triad. The inactive open conformation without a triad prevails in ligand-free solution and is converted to the closed conformation with a properly formed triad by the reaction product	Escherichia coli	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	-	Escherichia coli	(1R,6R)-2-succinyl-6-hydroxy-2,4-cyclohexadiene-1-carboxylate + pyruvate	-	?

Synonyms

Synonyms	Comment	Organism
MenH	-	Escherichia coli

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
0.13	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	mutant V152A, pH 7.0, temperature not specified in the publication	Escherichia coli
0.13	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	mutant Y148A, pH 7.0, temperature not specified in the publication	Escherichia coli
0.2	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	mutant V152G, pH 7.0, temperature not specified in the publication	Escherichia coli
0.58	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	mutant Y148F, pH 7.0, temperature not specified in the publication	Escherichia coli
3.33	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	mutant F153A, pH 7.0, temperature not specified in the publication	Escherichia coli
8.5	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	wild-type, pH 7.0, temperature not specified in the publication	Escherichia coli

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
0.43	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	mutant V152A, pH 7.0, temperature not specified in the publication	Escherichia coli
1.2	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	mutant V152G, pH 7.0, temperature not specified in the publication	Escherichia coli
2.7	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	mutant Y148A, pH 7.0, temperature not specified in the publication	Escherichia coli
16.3	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	mutant Y148F, pH 7.0, temperature not specified in the publication	Escherichia coli
48	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	mutant F153A, pH 7.0, temperature not specified in the publication	Escherichia coli
533	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	wild-type, pH 7.0, temperature not specified in the publication	Escherichia coli

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Release 2023.1 (January 2023)