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Literature summary extracted from

  • Odokonyero, D.; Ragumani, S.; Lopez, M.S.; Bonanno, J.B.; Ozerova, N.D.; Woodard, D.R.; Machala, B.W.; Swaminathan, S.; Burley, S.K.; Almo, S.C.; Glasner, M.E.
    Divergent evolution of ligand binding in the o-succinylbenzoate synthase family (2013), Biochemistry, 52, 7512-7521.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.2.1.113 gene menC, expression of N-terminally His-tagged wild-type and mutant enzymes in Escherichia coli strain BW25113 menC- Escherichia coli
4.2.1.113 gene menC, expression of N-terminally His-tagged wild-type and mutant enzymes in Escherichia coli strain BW25113 menC- Thermobifida fusca

Protein Variants

EC Number Protein Variants Comment Organism
4.2.1.113 F16A site-directed mutagenesis, the mutant enzyme activity is reduced compared to the wild-type enzyme Thermobifida fusca
4.2.1.113 F39A site-directed mutagenesis, the mutant enzyme activity is reduced compared to the wild-type enzyme Thermobifida fusca
4.2.1.113 G254A site-directed mutagenesis, the mutant enzyme activity is reduced compared to the wild-type enzyme Thermobifida fusca
4.2.1.113 G254T site-directed mutagenesis, the mutant enzyme activity is reduced compared to the wild-type enzyme Thermobifida fusca
4.2.1.113 G295A site-directed mutagenesis, the mutant enzyme activity is reduced compared to the wild-type enzyme Escherichia coli
4.2.1.113 N73A site-directed mutagenesis, the mutant enzyme activity is reduced compared to the wild-type enzyme Thermobifida fusca
4.2.1.113 N73A/T75A site-directed mutagenesis, the mutant is inactive Thermobifida fusca
4.2.1.113 N73A/T75L site-directed mutagenesis, the mutant enzyme activity is reduced compared to the wild-type enzyme Thermobifida fusca
4.2.1.113 N73L/T75V site-directed mutagenesis, the mutant enzyme activity is reduced compared to the wild-type enzyme Thermobifida fusca
4.2.1.113 R128M site-directed mutagenesis, the mutant enzyme activity is reduced compared to the wild-type enzyme Thermobifida fusca
4.2.1.113 R15G/F16G/R17G/G18G/I19G site-directed mutagenesis, the mutant enzyme activity is reduced compared to the wild-type enzyme Thermobifida fusca
4.2.1.113 R17A site-directed mutagenesis, the mutant enzyme activity is reduced compared to the wild-type enzyme Thermobifida fusca
4.2.1.113 R17A/F39A site-directed mutagenesis, the mutant enzyme activity is reduced compared to the wild-type enzyme Thermobifida fusca
4.2.1.113 R17A/Y42A site-directed mutagenesis, the mutant enzyme activity is reduced compared to the wild-type enzyme Thermobifida fusca
4.2.1.113 T75A site-directed mutagenesis, the mutant enzyme activity is reduced compared to the wild-type enzyme Thermobifida fusca
4.2.1.113 T75L site-directed mutagenesis, the mutant enzyme activity is reduced compared to the wild-type enzyme Thermobifida fusca
4.2.1.113 V230L site-directed mutagenesis, the mutant enzyme activity is reduced compared to the wild-type enzyme Thermobifida fusca
4.2.1.113 Y42A site-directed mutagenesis, the mutant enzyme activity is reduced compared to the wild-type enzyme Thermobifida fusca

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
4.2.1.113 0.007
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant R17A Thermobifida fusca
4.2.1.113 0.036
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant F16A Thermobifida fusca
4.2.1.113 0.038
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant R17A/Y42A Thermobifida fusca
4.2.1.113 0.048
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant R17A/F39A Thermobifida fusca
4.2.1.113 0.054
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant R15G/F16G/R17G/G18G/I19G Thermobifida fusca
4.2.1.113 0.057
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant T75L Thermobifida fusca
4.2.1.113 0.078
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant wild-type enzyme Escherichia coli
4.2.1.113 0.086
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant G254A Thermobifida fusca
4.2.1.113 0.145
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant V230L Thermobifida fusca
4.2.1.113 0.154
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant T75A Thermobifida fusca
4.2.1.113 0.182
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant wild-type enzyme Thermobifida fusca
4.2.1.113 0.256
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant N73A/T75L Thermobifida fusca
4.2.1.113 0.3
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant N73L/T75V Thermobifida fusca
4.2.1.113 0.304
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant G295A Escherichia coli
4.2.1.113 0.322
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant N73A Thermobifida fusca
4.2.1.113 0.47
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant R128M Thermobifida fusca
4.2.1.113 0.588
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant Y42A Thermobifida fusca
4.2.1.113 0.666
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant F39A Thermobifida fusca
4.2.1.113 0.668
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant G254T Thermobifida fusca

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
4.2.1.113 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate Escherichia coli
-
 2-succinylbenzoate + H2O
-
 ?
4.2.1.113 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate Thermobifida fusca
-
 2-succinylbenzoate + H2O
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
4.2.1.113 Escherichia coli P29208
-
-
4.2.1.113 Thermobifida fusca Q47Q21
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.2.1.113 recombinant N-terminally His-tagged wild-type and mutant enzymes from Escherichia coli strain BW25113 menC- by nickel affinity chromatography Escherichia coli
4.2.1.113 recombinant N-terminally His-tagged wild-type and mutant enzymes from Escherichia coli strain BW25113 menC- by nickel affinity chromatography Thermobifida fusca

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.2.1.113 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate
-
 Escherichia coli 2-succinylbenzoate + H2O
-
 ?
4.2.1.113 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate
-
 Thermobifida fusca 2-succinylbenzoate + H2O
-
 ?

Synonyms

EC Number Synonyms Comment Organism
4.2.1.113 OSBS
-
 Escherichia coli
4.2.1.113 OSBS
-
 Thermobifida fusca

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
4.2.1.113 25
-
 assay at Escherichia coli
4.2.1.113 25
-
 assay at Thermobifida fusca

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
4.2.1.113 0.1
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant R15G/F16G/R17G/G18G/I19G Thermobifida fusca
4.2.1.113 0.18
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant N73L/T75V Thermobifida fusca
4.2.1.113 0.2
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant R17A/F39A Thermobifida fusca
4.2.1.113 0.36
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant R17A/Y42A Thermobifida fusca
4.2.1.113 0.6
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant R17A Thermobifida fusca
4.2.1.113 4
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant G295A Escherichia coli
4.2.1.113 4.4
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant F16A Thermobifida fusca
4.2.1.113 5.1
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant N73A/T75L Thermobifida fusca
4.2.1.113 10
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant N73A Thermobifida fusca
4.2.1.113 13
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant G254T Thermobifida fusca
4.2.1.113 16
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant G254A Thermobifida fusca
4.2.1.113 18
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant T75L Thermobifida fusca
4.2.1.113 29
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant V230L Thermobifida fusca
4.2.1.113 55
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant T75A Thermobifida fusca
4.2.1.113 80
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant wild-type enzyme Escherichia coli
4.2.1.113 104
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant R128M Thermobifida fusca
4.2.1.113 122
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant wild-type enzyme Thermobifida fusca
4.2.1.113 163
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant F39A Thermobifida fusca
4.2.1.113 210
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant Y42A Thermobifida fusca

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
4.2.1.113 8
-
 assay at Escherichia coli
4.2.1.113 8
-
 assay at Thermobifida fusca

General Information

EC Number General Information Comment Organism
4.2.1.113 evolution o-succinylbenzoate synthase is a member of the enolase superfamily Escherichia coli
4.2.1.113 evolution o-succinylbenzoate synthase is a member of the enolase superfamily, there are significant differences in structure and conformational flexibility between Thermobifida fusca OSBS and other members of the enolase superfamily. The 20s loop, a flexible loop in the active site that permits ligand binding and release in most enolase superfamily proteins, has a four-amino acid deletion and is well-ordered in Thermobifida fusca OSBS. The flexibility of a different region allows the substrate to enter from the other side of the active site Thermobifida fusca
4.2.1.113 metabolism o-succinylbenzoate synthase catalyzes a step in menaquinone biosynthesis Escherichia coli
4.2.1.113 metabolism o-succinylbenzoate synthase catalyzes a step in menaquinone biosynthesis Thermobifida fusca
4.2.1.113 additional information the 20s loop, a flexible loop in the active site that permits ligand binding and release in most enolase superfamily proteins, has a four-amino acid deletion and is well-ordered in Thermobifida fusca OSBS. The flexibility of a different region allows the substrate to enter from the other side of the active site, active site structure, overview. Comparison of apo and ligand-bound structures of the Mycobacterium tuberculosis enzyme with the Escherichia coli enzyme Thermobifida fusca

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
4.2.1.113 0.61
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant N73L/T75V Thermobifida fusca
4.2.1.113 4.2
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant R17A/F39A Thermobifida fusca
4.2.1.113 9.5
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant R17A/Y42A Thermobifida fusca
4.2.1.113 13
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant G295A Escherichia coli
4.2.1.113 19
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant G254T Thermobifida fusca
4.2.1.113 20
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant N73A/T75L Thermobifida fusca
4.2.1.113 31
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant N73A Thermobifida fusca
4.2.1.113 86
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant R17A Thermobifida fusca
4.2.1.113 120
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant F16A Thermobifida fusca
4.2.1.113 122
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant wild-type enzyme Thermobifida fusca
4.2.1.113 190
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant G254A Thermobifida fusca
4.2.1.113 200
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant V230L Thermobifida fusca
4.2.1.113 220
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant R128M Thermobifida fusca
4.2.1.113 250
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant F39A Thermobifida fusca
4.2.1.113 320
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant T75L Thermobifida fusca
4.2.1.113 360
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant T75A Thermobifida fusca
4.2.1.113 360
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant Y42A Thermobifida fusca
4.2.1.113 670
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant mutant R15G/F16G/R17G/G18G/I19G Thermobifida fusca
4.2.1.113 1000
-
 (1R,6R)-6-hydroxy-2-succinylcyclohexa-2,4-diene-1-carboxylate pH 8.0, 25°C, recombinant wild-type enzyme Escherichia coli