Literature summary for 4.2.99.20 extracted from

Jiang, M.; Chen, X.; Wu, X.H.; Chen, M.; Wu, Y.D.; Guo, Z.
Catalytic mechanism of SHCHC synthase in the menaquinone biosynthesis of Escherichia coli: identification and mutational analysis of the active site residues (2009), Biochemistry, 48, 6921-6931.

Search for more literature for 4.2.99.20

Cloned(Commentary)

Cloned (Comment)	Organism
expressed in Escherichia coli BL21(DE3)	Escherichia coli

Protein Variants

Protein Variants	Comment	Organism
D128A	conserved among MenH proteins	Escherichia coli
D210A	part of the catalytically essential triad	Escherichia coli
H232A	part of the catalytically essential triad	Escherichia coli
H232K	part of the catalytically essential triad	Escherichia coli
K212A	conserved among MenH proteins	Escherichia coli
R124A	conserved among MenH proteins	Escherichia coli
R168A	conserved among MenH proteins	Escherichia coli
R90A	conserved among MenH proteins	Escherichia coli
S86A	part of the catalytically essential triad	Escherichia coli
W147A	conserved among MenH proteins	Escherichia coli
W147F	conserved among MenH proteins	Escherichia coli
Y85F	conserved among MenH proteins	Escherichia coli

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.0086	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	H232A mutant protein	Escherichia coli
0.01	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	wild-type protein	Escherichia coli
0.014	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	Y85F mutant protein	Escherichia coli
0.0194	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	K212A mutant protein	Escherichia coli
0.0237	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	W147A mutant protein	Escherichia coli
0.028	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	S86A mutant protein	Escherichia coli
0.074	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	W147F mutant protein	Escherichia coli
0.084	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	R124A mutant protein	Escherichia coli
0.085	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	D128A mutant protein	Escherichia coli
0.118	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	D210A mutant protein	Escherichia coli
0.134	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	R168A mutant protein	Escherichia coli
0.159	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	R90A mutant protein	Escherichia coli
0.532	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	H232K mutant protein	Escherichia coli

Organism

Organism	UniProt	Comment	Textmining
Escherichia coli	-	-	-

Purification (Commentary)

Purification (Comment)	Organism
immobilized metal ion affinity chromatography, gel filtration	Escherichia coli

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
(1R,6R)-2-succinyl-6-hydroxy-2,4-cyclohexadiene-1-carboxylate synthase	-	Escherichia coli
MenH	-	Escherichia coli
SHCHC	-	Escherichia coli

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
0.00058	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	H232A mutant protein	Escherichia coli
0.021	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	H232K mutant protein	Escherichia coli
0.047	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	R90A mutant protein	Escherichia coli
0.056	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	W147A mutant protein	Escherichia coli
0.18	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	D210A mutant protein	Escherichia coli
0.29	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	S86A mutant protein	Escherichia coli
0.51	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	R124A mutant protein	Escherichia coli
2.58	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	Y85F mutant protein	Escherichia coli
2.9	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	R168A mutant protein	Escherichia coli
15.4	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	W147F mutant protein	Escherichia coli
41.5	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	K212A mutant protein	Escherichia coli
115	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	D128A mutant protein	Escherichia coli
147	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	wild-type protein	Escherichia coli

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
0.0392	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	H232K mutant protein	Escherichia coli
0.067	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	H232A mutant protein	Escherichia coli
0.295	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	R90A mutant protein	Escherichia coli
1.5	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	D210A mutant protein	Escherichia coli
2.4	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	W147A mutant protein	Escherichia coli
6.1	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	R124A mutant protein	Escherichia coli
10	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	S86A mutant protein	Escherichia coli
15	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	R168A mutant protein	Escherichia coli
180	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	Y85F mutant protein	Escherichia coli
220	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	W147F mutant protein	Escherichia coli
1400	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	D128A mutant protein	Escherichia coli
2200	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	K212A mutant protein	Escherichia coli
14000	-	(1R,2S,5S,6S)-2-succinyl-5-enolpyruvyl-6-hydroxy-3-cyclohexene-1-carboxylate	wild-type protein	Escherichia coli

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Release 2023.1 (January 2023)