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Literature summary 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)

EC Number Cloned (Comment) Organism
6.3.2.17 expression in Escherichia coli Lacticaseibacillus casei

Protein Variants

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

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
6.3.2.17 0.0315
-
 5,6,7,8-tetrahydrofolic acid pH 7.5, wild type enzyme Lacticaseibacillus casei
6.3.2.17 0.071
-
 5,6,7,8-tetrahydrofolic acid pH 7.5, E81A mutant Lacticaseibacillus casei
6.3.2.17 0.094
-
 5,6,7,8-tetrahydrofolic acid pH 7.5, S412A mutant Lacticaseibacillus casei
6.3.2.17 0.21
-
 5,6,7,8-tetrahydrofolic acid pH 7.5, R82A mutant Lacticaseibacillus casei
6.3.2.17 0.24
-
 5,6,7,8-tetrahydrofolic acid pH 7.5, K185A mutant Lacticaseibacillus casei
6.3.2.17 0.41
-
 L-glutamate pH 7.5, E81A mutant Lacticaseibacillus casei
6.3.2.17 0.47
-
 L-glutamate pH 7.5, wild type enzyme Lacticaseibacillus casei
6.3.2.17 0.62
-
 ATP pH 7.5, S412A mutant Lacticaseibacillus casei
6.3.2.17 0.655
-
 5,6,7,8-tetrahydrofolic acid pH 7.5, S73A mutant Lacticaseibacillus casei
6.3.2.17 0.85
-
 L-glutamate pH 7.5, S73A mutant Lacticaseibacillus casei
6.3.2.17 0.87
-
 ATP pH 7.5, R82A mutant Lacticaseibacillus casei
6.3.2.17 2.06
-
 L-glutamate pH 7.5, R82A mutant Lacticaseibacillus casei
6.3.2.17 2.5
-
 ATP pH 7.5, S73A mutant Lacticaseibacillus casei
6.3.2.17 2.9
-
 ATP pH 7.5, K185A mutant Lacticaseibacillus casei
6.3.2.17 3.4
-
 ATP pH 7.5, wild type enzyme Lacticaseibacillus casei
6.3.2.17 5.06
-
 L-glutamate pH 7.5, K185A mutant Lacticaseibacillus casei
6.3.2.17 5.2
-
 ATP pH 7.5, E81A mutant Lacticaseibacillus casei
6.3.2.17 26.69
-
 L-glutamate pH 7.5, S412A mutant Lacticaseibacillus casei

Organism

EC Number Organism UniProt Comment Textmining
6.3.2.17 Lacticaseibacillus casei P15925
-
-

Purification (Commentary)

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

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
6.3.2.17 0.0015
-
 K185A mutant Lacticaseibacillus casei
6.3.2.17 0.0043
-
 S412A mutant Lacticaseibacillus casei
6.3.2.17 0.0053
-
 E81A mutant Lacticaseibacillus casei
6.3.2.17 0.008
-
 R82A mutant Lacticaseibacillus casei
6.3.2.17 0.0093
-
 S73A mutant Lacticaseibacillus casei
6.3.2.17 0.2
-
 wild type enzyme Lacticaseibacillus casei

Storage Stability

EC Number Storage Stability Organism
6.3.2.17 -70ºC, 20% DMSO, stable Lacticaseibacillus casei

Substrates and Products (Substrate)

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