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Literature summary extracted from

  • Patin, D.; Turk, S.; Barreteau, H.; Mainardi, J.L.; Arthur, M.; Gobec, S.; Mengin-Lecreulx, D.; Blanot, D.
    Unusual substrate specificity of the peptidoglycan MurE ligase from Erysipelothrix rhusiopathiae (2016), Biochimie, 121, 209-218 .
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
6.3.2.13 gene murE, recombinant overexpression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)/pLysS, subcloning in Escherichia coli strain DH5alpha Erysipelothrix rhusiopathiae

Protein Variants

EC Number Protein Variants Comment Organism
6.3.2.13 D399A site-directed mutagenesis, inactive mutant Erysipelothrix rhusiopathiae
6.3.2.13 D399N site-directed mutagenesis, almost inactive mutant Erysipelothrix rhusiopathiae
6.3.2.13 F340A site-directed mutagenesis, almost inactive mutant Erysipelothrix rhusiopathiae
6.3.2.13 F340Y site-directed mutagenesis, the mutant shows reduced activity with substrate L-alanine and meso-2,6-diaminoheptanedioate compared to wild-type enzyme Erysipelothrix rhusiopathiae
6.3.2.13 H398A site-directed mutagenesis, the mutant shows reduced activity with substrate L-alanine and meso-2,6-diaminoheptanedioate compared to wild-type enzyme Erysipelothrix rhusiopathiae
6.3.2.13 H398D site-directed mutagenesis, the mutant shows highly reduced activity with substrate L-alanine and meso-2,6-diaminoheptanedioate compared to wild-type enzyme Erysipelothrix rhusiopathiae
6.3.2.13 additional information construction of an enzyme mutant MurEEr(HDNR/DNPR) with exchange of the HDNR morif to a DNPR motif, which is characteristic of meso-diaminopimelate-adding enzymes. The mutant shows increased activity with substrate meso-2,6-diaminoheptanedioate and reduced activity with substrate L-alanine Erysipelothrix rhusiopathiae
6.3.2.13 N400A site-directed mutagenesis, the mutant shows reduced activity with substrate L-alanine and meso-2,6-diaminoheptanedioate compared to wild-type enzyme Erysipelothrix rhusiopathiae
6.3.2.13 N400P site-directed mutagenesis, the mutant shows slightly reduced activity with substrate meso-2,6-diaminoheptanedioate, but increased activity with substrate L-alanine compared to wild-type enzyme Erysipelothrix rhusiopathiae
6.3.2.13 R401A site-directed mutagenesis, almost inactive mutant Erysipelothrix rhusiopathiae

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
6.3.2.13 0.16
-
 ATP pH 9.6, 37°C, recombinant His-tagged enzyme Erysipelothrix rhusiopathiae
6.3.2.13 0.16
-
 UDP-N-acetyl-alpha-D-muramoyl-L-alanyl-D-glutamate pH 9.6, 37°C, recombinant His-tagged enzyme Erysipelothrix rhusiopathiae
6.3.2.13 0.21
-
 UDP-N-acetyl-alpha-D-muramoyl-L-seryl-D-glutamate pH 9.6, 37°C, recombinant His-tagged enzyme Erysipelothrix rhusiopathiae
6.3.2.13 0.23
-
 UDP-N-acetyl-alpha-D-muramoyl-glycyl-D-glutamate pH 9.6, 37°C, recombinant His-tagged enzyme Erysipelothrix rhusiopathiae
6.3.2.13 1.8
-
 L-alanine pH 9.6, 37°C, recombinant His-tagged enzyme Erysipelothrix rhusiopathiae
6.3.2.13 10
-
 meso-2,6-diaminoheptanedioate pH 9.6, 37°C, recombinant mutant MurEEr(HDNR/DNPR) Erysipelothrix rhusiopathiae
6.3.2.13 64
-
 L-alanine pH 9.6, 37°C, recombinant mutant MurEEr(HDNR/DNPR) Erysipelothrix rhusiopathiae
6.3.2.13 77
-
 meso-2,6-diaminoheptanedioate pH 9.6, 37°C, recombinant His-tagged enzyme Erysipelothrix rhusiopathiae

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
6.3.2.13 Mg2+ required Erysipelothrix rhusiopathiae

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
6.3.2.13 ATP + UDP-N-acetyl-alpha-D-muramoyl-L-alanyl-D-glutamate + meso-2,6-diaminoheptanedioate Erysipelothrix rhusiopathiae
-
 ADP + phosphate + UDP-N-acetyl-alpha-D-muramoyl-L-alanyl-gamma-D-glutamyl-meso-2,6-diaminoheptanedioate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
6.3.2.13 Erysipelothrix rhusiopathiae E7FUC9
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
6.3.2.13 recombinant His-tagged wild-type and mutant enzyme from Escherichia coli strain BL21(DE3)/pLysS by nickel affinity chromatography Erysipelothrix rhusiopathiae

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
6.3.2.13 ATP + UDP-N-acetyl-alpha-D-muramoyl-glycyl-D-glutamate + L-alanine
-
 Erysipelothrix rhusiopathiae ADP + phosphate + UDP-N-acetyl-alpha-D-muramoyl-glycyl-gamma-D-glutamyl-L-alanine
-
 ?
6.3.2.13 ATP + UDP-N-acetyl-alpha-D-muramoyl-L-alanyl-D-glutamate + (2S,6S)-2,6-diaminoheptanedioate 7.8% activity with (2S,6S)-2,6-diaminoheptanedioate compared to L-alanine Erysipelothrix rhusiopathiae ADP + phosphate + UDP-N-acetyl-alpha-D-muramoyl-L-alanyl-(2S,6S)-2,6-diaminoheptanedioate
-
 ?
6.3.2.13 ATP + UDP-N-acetyl-alpha-D-muramoyl-L-alanyl-D-glutamate + beta-chloro-L-alanine 15% activity with beta-chloro-L-alanine compared to L-alanine Erysipelothrix rhusiopathiae ADP + phosphate + UDP-N-acetyl-alpha-D-muramoyl-L-alanyl-beta-chloro-L-alanine
-
 ?
6.3.2.13 ATP + UDP-N-acetyl-alpha-D-muramoyl-L-alanyl-D-glutamate + glycine 7% activity with glycine compared to L-alanine Erysipelothrix rhusiopathiae ADP + phosphate + UDP-N-acetyl-alpha-D-muramoyl-L-alanyl-glycine
-
 ?
6.3.2.13 ATP + UDP-N-acetyl-alpha-D-muramoyl-L-alanyl-D-glutamate + L-alanine highest activity with L-alanine compared to the other amino acids Erysipelothrix rhusiopathiae ADP + phosphate + UDP-N-acetyl-alpha-D-muramoyl-L-alanyl-gamma-D-glutamyl-L-alanine
-
 ?
6.3.2.13 ATP + UDP-N-acetyl-alpha-D-muramoyl-L-alanyl-D-glutamate + L-diaminobutyrate 7.9% activity with L-diaminobutyrate compared to L-alanine Erysipelothrix rhusiopathiae ADP + phosphate + UDP-N-acetyl-alpha-D-muramoyl-L-alanyl-L-diaminobutyrate
-
 ?
6.3.2.13 ATP + UDP-N-acetyl-alpha-D-muramoyl-L-alanyl-D-glutamate + L-homoserine 10% activity with L-homoserine compared to L-alanine Erysipelothrix rhusiopathiae ADP + phosphate + UDP-N-acetyl-alpha-D-muramoyl-L-alanyl-L-homoserine
-
 ?
6.3.2.13 ATP + UDP-N-acetyl-alpha-D-muramoyl-L-alanyl-D-glutamate + L-propargylglycine 20% activity with L-propargylglycine compared to L-alanine Erysipelothrix rhusiopathiae ADP + phosphate + UDP-N-acetyl-alpha-D-muramoyl-L-alanyl-L-propargylglycine
-
 ?
6.3.2.13 ATP + UDP-N-acetyl-alpha-D-muramoyl-L-alanyl-D-glutamate + L-serine 30% activity with L-serine compared to L-alanine Erysipelothrix rhusiopathiae ADP + phosphate + UDP-N-acetyl-alpha-D-muramoyl-L-alanyl-L-serine
-
 ?
6.3.2.13 ATP + UDP-N-acetyl-alpha-D-muramoyl-L-alanyl-D-glutamate + L-vinylglycine 58% activity with L-vinylglycine compared to L-alanine Erysipelothrix rhusiopathiae ADP + phosphate + UDP-N-acetyl-alpha-D-muramoyl-L-alanyl-L-vinylglycine
-
 ?
6.3.2.13 ATP + UDP-N-acetyl-alpha-D-muramoyl-L-alanyl-D-glutamate + meso-2,6-diaminoheptanedioate
-
 Erysipelothrix rhusiopathiae ADP + phosphate + UDP-N-acetyl-alpha-D-muramoyl-L-alanyl-gamma-D-glutamyl-meso-2,6-diaminoheptanedioate
-
 ?
6.3.2.13 ATP + UDP-N-acetyl-alpha-D-muramoyl-L-alanyl-D-glutamate + meso-2,6-diaminoheptanedioate 30% activity with meso-2,6-diaminoheptanedioate compared to L-alanine Erysipelothrix rhusiopathiae ADP + phosphate + UDP-N-acetyl-alpha-D-muramoyl-L-alanyl-gamma-D-glutamyl-meso-2,6-diaminoheptanedioate
-
 ?
6.3.2.13 ATP + UDP-N-acetyl-alpha-D-muramoyl-L-seryl-D-glutamate + L-alanine
-
 Erysipelothrix rhusiopathiae ADP + phosphate + UDP-N-acetyl-alpha-D-muramoyl-L-seryl-gamma-D-glutamyl-L-alanine
-
 ?
6.3.2.13 additional information enzyme substrate specificity, overview. Poor activity with D-alanine, L-lysine, L-ornithine, L-cysteine, L-allylglycine, L-1-aminoethylphosphonic acid, L-alanine amide, L-norvaline and L-alpha-aminobutyric acid. Enzyme-ligand interaction analysis, overview Erysipelothrix rhusiopathiae ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
6.3.2.13 Meso-diaminopimelate-adding enzyme
-
 Erysipelothrix rhusiopathiae
6.3.2.13 MurE
-
 Erysipelothrix rhusiopathiae
6.3.2.13 MurEEr
-
 Erysipelothrix rhusiopathiae
6.3.2.13 peptidoglycan MurE ligase
-
 Erysipelothrix rhusiopathiae

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
6.3.2.13 37
-
 assay at Erysipelothrix rhusiopathiae

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
6.3.2.13 1.03
-
 L-alanine pH 9.6, 37°C, recombinant mutant MurEEr(HDNR/DNPR) Erysipelothrix rhusiopathiae
6.3.2.13 1.83
-
 ATP pH 9.6, 37°C, recombinant His-tagged enzyme Erysipelothrix rhusiopathiae
6.3.2.13 2.83
-
 L-alanine pH 9.6, 37°C, recombinant His-tagged enzyme Erysipelothrix rhusiopathiae
6.3.2.13 3.33
-
 UDP-N-acetyl-alpha-D-muramoyl-L-alanyl-D-glutamate pH 9.6, 37°C, recombinant His-tagged enzyme Erysipelothrix rhusiopathiae
6.3.2.13 3.67
-
 UDP-N-acetyl-alpha-D-muramoyl-L-seryl-D-glutamate pH 9.6, 37°C, recombinant His-tagged enzyme Erysipelothrix rhusiopathiae
6.3.2.13 4.17
-
 meso-2,6-diaminoheptanedioate pH 9.6, 37°C, recombinant His-tagged enzyme Erysipelothrix rhusiopathiae
6.3.2.13 4.83
-
 meso-2,6-diaminoheptanedioate pH 9.6, 37°C, recombinant mutant MurEEr(HDNR/DNPR) Erysipelothrix rhusiopathiae
6.3.2.13 8.83
-
 UDP-N-acetyl-alpha-D-muramoyl-glycyl-D-glutamate pH 9.6, 37°C, recombinant His-tagged enzyme Erysipelothrix rhusiopathiae

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
6.3.2.13 9.6
-
 assay at Erysipelothrix rhusiopathiae

Cofactor

EC Number Cofactor Comment Organism Structure
6.3.2.13 ATP
-
 Erysipelothrix rhusiopathiae

General Information

EC Number General Information Comment Organism
6.3.2.13 evolution the DNPR motif that is characteristic of meso-diaminopimelate-adding enzymes is replaced by HDNR Erysipelothrix rhusiopathiae
6.3.2.13 additional information the HDNR motif plays a role in the interaction of the enzyme with L-alanine and meso-diaminopimelic acid. ENzyme structure homology modeling Erysipelothrix rhusiopathiae

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
6.3.2.13 0.016
-
 L-alanine pH 9.6, 37°C, recombinant mutant MurEEr(HDNR/DNPR) Erysipelothrix rhusiopathiae
6.3.2.13 0.054
-
 meso-2,6-diaminoheptanedioate pH 9.6, 37°C, recombinant His-tagged enzyme Erysipelothrix rhusiopathiae
6.3.2.13 0.483
-
 meso-2,6-diaminoheptanedioate pH 9.6, 37°C, recombinant mutant MurEEr(HDNR/DNPR) Erysipelothrix rhusiopathiae
6.3.2.13 1.57
-
 L-alanine pH 9.6, 37°C, recombinant His-tagged enzyme Erysipelothrix rhusiopathiae
6.3.2.13 11.44
-
 ATP pH 9.6, 37°C, recombinant His-tagged enzyme Erysipelothrix rhusiopathiae
6.3.2.13 17.48
-
 UDP-N-acetyl-alpha-D-muramoyl-L-seryl-D-glutamate pH 9.6, 37°C, recombinant His-tagged enzyme Erysipelothrix rhusiopathiae
6.3.2.13 20.81
-
 UDP-N-acetyl-alpha-D-muramoyl-L-alanyl-D-glutamate pH 9.6, 37°C, recombinant His-tagged enzyme Erysipelothrix rhusiopathiae
6.3.2.13 38.39
-
 UDP-N-acetyl-alpha-D-muramoyl-glycyl-D-glutamate pH 9.6, 37°C, recombinant His-tagged enzyme Erysipelothrix rhusiopathiae