Literature summary for 2.5.1.19 extracted from

Funke, T.; Yang, Y.; Han, H.; Healy-Fried, M.; Olesen, S.; Becker, A.; Schoenbrunn, E.
Structural basis of glyphosate resistance resulting from the double mutation Thr97 Ile and Pro101 Ser in 5-enolpyruvylshikimate-3-phosphate synthase from Escherichia coli (2009), J. Biol. Chem., 284, 9854-9860.

Search for more literature for 2.5.1.19

Cloned(Commentary)

Cloned (Comment)	Organism
overexpressed in Escherichia coli	Escherichia coli

Crystallization (Commentary)

Crystallization (Comment)	Organism
using the hanging drop, vapor-diffusion method in the presence of 5 mM 3-phosphoshikimate	Escherichia coli

Protein Variants

Protein Variants	Comment	Organism
P101S	the substrate binding affinities, as reflected by the respective Km values, are only slightly decreased for the P101S mutant. Km (3-phosphoshikimate): 0.071 mM, Km (phosphoenolpyruvate): 0.071, kcat/Km (phosphoenolpyruvate): 230000/Msec, kcat/Km (3-phosphoshikimate): 240000/Msec. Mutant P101S is moderately inhibited by glyphosate	Escherichia coli
T971I	the single site T97I mutation renders the enzyme sensitive to glyphosate and causes a substantial decrease in the affinity for phosphoenolpyruvate. Km (3-phosphoshikimate): 0.077 mM, Km (phosphoenolpyruvate): 0.38, kcat/Km (phosphoenolpyruvate): 23000/Msec, kcat/Km (3-phosphoshikimate): 1200000/Msec	Escherichia coli
T97I/P101S	mutant is essentially insensitive to glyphosate (Ki 2.4 mM) but maintains high affinity for the substrate phosphoenolpyruvate (Km: 0.1 mM) and 3-phosphoshikimate (Km: 0.077 mM). kcat/Km (phosphoenolpyruvate): 57000/Msec, kcat/Km (3-phosphoshikimate): 740000/Msec. The crystal structure at 1.7 A resolution reveals that the dual mutation causes a shift of residue Gly96 toward the glyphosate binding site, impairing efficient binding of glyphosate, while the side chain of Ile97 points away from the substrate binding site, facilitating phosphoenolpyruvate utilization	Escherichia coli

Inhibitors

Inhibitors	Comment	Organism	Structure
glyphosate	-	Escherichia coli

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.045	-	phosphoenolpyruvate	wild-type	Escherichia coli
0.048	-	3-phosphoshikimate	wild-type	Escherichia coli
0.071	-	phosphoenolpyruvate	mutant P101S	Escherichia coli
0.071	-	3-phosphoshikimate	mutant P101S	Escherichia coli
0.077	-	3-phosphoshikimate	mutant P101S/T97I	Escherichia coli
0.077	-	3-phosphoshikimate	mutant T97I	Escherichia coli
0.1	-	phosphoenolpyruvate	mutant P101S/T97I	Escherichia coli
0.38	-	phosphoenolpyruvate	mutant T97I	Escherichia coli

Organism

Organism	UniProt	Comment	Textmining
Escherichia coli	P0A6D3	-	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
phosphoenolpyruvate + 3-phosphoshikimate	-	Escherichia coli	phosphate + 5-enolpyruvylshikimate 3-phosphate	-	?

Synonyms

Synonyms	Comment	Organism
EPSPS	-	Escherichia coli

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.5	-	assay at	Escherichia coli

Ki Value [mM]

Ki Value [mM]	Ki Value maximum [mM]	Inhibitor	Comment	Organism	Structure
0.0003	-	glyphosate	wild-type	Escherichia coli
0.003	-	glyphosate	mutant P101S	Escherichia coli
0.09	-	glyphosate	mutant P101S	Escherichia coli

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
23000000	-	phosphoenolpyruvate	mutant T97I	Escherichia coli
57000000	-	phosphoenolpyruvate	mutant P101S/T97I	Escherichia coli
74000000	-	3-phosphoshikimate	mutant P101S/T97I	Escherichia coli
120000000	-	3-phosphoshikimate	mutant T97I	Escherichia coli
240000000	-	3-phosphoshikimate	mutant P101S	Escherichia coli
250000000	-	phosphoenolpyruvate	mutant P101S	Escherichia coli
910000000	-	3-phosphoshikimate	wild-type	Escherichia coli
930000000	-	phosphoenolpyruvate	wild-type	Escherichia coli

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Release 2023.1 (January 2023)