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Literature summary for 6.3.2.17 extracted from

  • Sheng, Y.; Sun, X.; Shen, Y.; Bognar, A.L.; Baker, E.N.; Smith, C.A.
    Structural and functional similarities in the ADP-forming amide bond ligase superfamily: implications for a substrate-induced conformational change in folylpolyglutamate synthetase (2000), J. Mol. Biol., 302, 427-440.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Lacticaseibacillus casei

Protein Variants

Protein Variants Comment Organism
D313A mutation in the C-terminal of the enzyme Lacticaseibacillus casei
E143A mutation in the MgATP2- binding site of the enzyme Lacticaseibacillus casei
E143D mutation in the MgATP2- binding site of the enzyme Lacticaseibacillus casei
E143Q mutation in the MgATP2- binding site of the enzyme Lacticaseibacillus casei
E81A mutation in the omega-loop of the enzyme Lacticaseibacillus casei
G411A mutation in the C-terminal of the enzyme Lacticaseibacillus casei
H316A mutation in the C-terminal of the enzyme Lacticaseibacillus casei
K185A mutation in the MgATP2- binding site of the enzyme Lacticaseibacillus casei
P74A mutation in the omega-loop of the enzyme Lacticaseibacillus casei
R82A mutation in the omega-loop of the enzyme Lacticaseibacillus casei
S412A mutation in the C-terminal of the enzyme Lacticaseibacillus casei
S73A mutation in the omega-loop of the enzyme Lacticaseibacillus casei

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0315
-
5,6,7,8-tetrahydrofolic acid pH 7.5, wild type enzyme Lacticaseibacillus casei
0.071
-
5,6,7,8-tetrahydrofolic acid pH 7.5, E81A mutant Lacticaseibacillus casei
0.094
-
5,6,7,8-tetrahydrofolic acid pH 7.5, S412A mutant Lacticaseibacillus casei
0.21
-
5,6,7,8-tetrahydrofolic acid pH 7.5, R82A mutant Lacticaseibacillus casei
0.24
-
5,6,7,8-tetrahydrofolic acid pH 7.5, K185A mutant Lacticaseibacillus casei
0.41
-
L-glutamate pH 7.5, E81A mutant Lacticaseibacillus casei
0.47
-
L-glutamate pH 7.5, wild type enzyme Lacticaseibacillus casei
0.62
-
ATP pH 7.5, S412A mutant Lacticaseibacillus casei
0.655
-
5,6,7,8-tetrahydrofolic acid pH 7.5, S73A mutant Lacticaseibacillus casei
0.85
-
L-glutamate pH 7.5, S73A mutant Lacticaseibacillus casei
0.87
-
ATP pH 7.5, R82A mutant Lacticaseibacillus casei
2.06
-
L-glutamate pH 7.5, R82A mutant Lacticaseibacillus casei
2.5
-
ATP pH 7.5, S73A mutant Lacticaseibacillus casei
2.9
-
ATP pH 7.5, K185A mutant Lacticaseibacillus casei
3.4
-
ATP pH 7.5, wild type enzyme Lacticaseibacillus casei
5.06
-
L-glutamate pH 7.5, K185A mutant Lacticaseibacillus casei
5.2
-
ATP pH 7.5, E81A mutant Lacticaseibacillus casei
26.69
-
L-glutamate pH 7.5, S412A mutant Lacticaseibacillus casei

Organism

Organism UniProt Comment Textmining
Lacticaseibacillus casei P15925
-
-

Purification (Commentary)

Purification (Comment) Organism
chromatography on a chitin column Lacticaseibacillus casei

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
0.0015
-
K185A mutant Lacticaseibacillus casei
0.0043
-
S412A mutant Lacticaseibacillus casei
0.0053
-
E81A mutant Lacticaseibacillus casei
0.008
-
R82A mutant Lacticaseibacillus casei
0.0093
-
S73A mutant Lacticaseibacillus casei
0.2
-
wild type enzyme Lacticaseibacillus casei

Storage Stability

Storage Stability Organism
-70ºC, 20% DMSO, stable Lacticaseibacillus casei

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + 5,6,7,8-tetrahydrofolic acid + L-glutamate
-
Lacticaseibacillus casei ADP + phosphate + 5,6,7,8-tetrahydrofolyl-Glu
-
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