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Literature summary extracted from

  • Park, I.-S.; Lin, C.-H.; Walsh, C.T.
    Gain of D-alanyl-D-lactate or D-lactyl-D-alanine synthetase activities in three active-site mutants of the Escherichia coli D-alanyl-D-alanine ligase B (1996), Biochemistry, 35, 10464-10471.
    View publication on PubMed

Protein Variants

EC Number Protein Variants Comment Organism
6.1.2.1 S150A mutant of D-Ala-D-Ala ligase Ddl, mutant has gained depsipeptide ligase activity, i.e. formation of D-Ala-D-Lac, D-Ala-D-hydroxybutyrate, with dipeptide/depsipeptide partition ratios that mimic the pH behaviour of D-Ala-D-lactate ligase VanA. Mutant displays a clear pH-dependent partitioning between the preferred depsipeptide product at low pH and the dipeptide product at high pH Escherichia coli
6.1.2.1 Y216F mutant of D-Ala-D-Ala ligase Ddl, mutant has gained depsipeptide ligase activity, i.e. formation of D-Ala-D-Lac, D-Ala-D-hydroxybutyrate, with dipeptide/depsipeptide partition ratios that mimic the pH behaviour of D-Ala-D-lactate ligase VanA. Mutant displays a clear pH-dependent partitioning between the preferred depsipeptide product at low pH and the dipeptide product at high pH Escherichia coli
6.3.2.4 E15Q mutant enzyme with negligible depsipeptide synthetase activity. The mutant uniquely activates D-Lac as the electrophilic rather than the nucleophilic partner for condensation with D-Ala to make a regioisomeric D-Lac-D-Ala, an amide rather than an ester product Escherichia coli
6.3.2.4 S150A mutant has gained depsipeptide (D-Ala-D-Lac, D-Ala-D-hydroxybutyrate) ligase activity with dipeptide/depsipeptide partition ratios that mimic the pH behavior of VanA (D-alanine—(R)-lactate ligase) Escherichia coli
6.3.2.4 Y216F mutant has gained depsipeptide (D-Ala-D-Lac, D-Ala-D-hydroxybutyrate) ligase activity with dipeptide/depsipeptide partition ratios that mimic the pH behavior of VanA (D-alanine—(R)-lactate ligase) Escherichia coli

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
6.1.2.1 0.88
-
 D-lactate isoform VanA, pH 7.5, 37°C Escherichia coli
6.1.2.1 1.3
-
 D-lactate isoform VanA, pH 6.0, 37°C Escherichia coli
6.1.2.1 1.5
-
 D-lactate isoform VanA, pH 8.3, 37°C Escherichia coli
6.1.2.1 1.6
-
 D-lactate mutant S150A, pH 6.0, 37°C Escherichia coli
6.1.2.1 2.2
-
 D-lactate mutant S150A, pH 7.5, 37°C Escherichia coli
6.1.2.1 5.1
-
 D-lactate mutant Y216F, pH 6.0, 37°C Escherichia coli
6.1.2.1 16
-
 D-Ala mutant Y216F, pH 7.5, 37°C Escherichia coli
6.1.2.1 27
-
 D-lactate mutant Y216F, pH 7.5, 37°C Escherichia coli
6.1.2.1 69
-
 D-Ala isoform VanA, pH 8.3, 37°C Escherichia coli
6.1.2.1 80
-
 D-Ala mutant S150A, pH 7.5, 37°C Escherichia coli
6.1.2.1 100
-
 D-Ala isoform VanA, above 100 mM, pH 7.5, 37°C Escherichia coli
6.1.2.1 140
-
 D-Ala mutant S150A, pH 6.0, 37°C Escherichia coli
6.1.2.1 150
-
 D-Ala mutant Y216F, pH 6.0, 37°C Escherichia coli
6.3.2.4 0.003
-
 D-alanine 37°C, KM-value for D-alanine1 (the first molecule of substrate that is activated by the enzyme molecule) , wild-type enzyme, pH 7.5 Escherichia coli
6.3.2.4 0.004
-
 D-alanine 37°C, KM-value for D-alanine1 (the first molecule of substrate that is activated by the enzyme molecule), wild-type enzyme, pH 9.2 Escherichia coli
6.3.2.4 0.11
-
 D-alanine 37°C, KM-value for D-alanine1 (the first molecule of substrate that is activated by the enzyme molecule), wild-type enzyme, pH 6.0 Escherichia coli
6.3.2.4 0.51
-
 D-alanine 37°C, KM-value for D-alanine1 (the first molecule of substrate that is activated by the enzyme molecule), mutant enzyme E15Q, pH 7.5 Escherichia coli
6.3.2.4 0.51
-
 D-alanine 37°C, KM-value for D-alanine1 (the first molecule of substrate that is activated by the enzyme molecule), mutant enzyme E15Q, pH 9.2 Escherichia coli
6.3.2.4 0.85
-
 D-alanine 37°C, KM-value for D-alanine1 (the first molecule of substrate that is activated by the enzyme molecule), mutant enzyme S150A, pH 7.5 Escherichia coli
6.3.2.4 1.1
-
 D-alanine 37°C, KM-value for D-alanine2 (the second molecule of substrate that is activated by the enzyme molecule), wild-type enzyme, pH 9.2 Escherichia coli
6.3.2.4 2
-
 D-alanine 37°C, KM-value for D-alanine2 (the second molecule of substrate that is activated by the enzyme molecule), wild-type enzyme, pH 7.5 Escherichia coli
6.3.2.4 4.8
-
 D-alanine 37°C, KM-value for D-alanine1 (the first molecule of substrate that is activated by the enzyme molecule), mutant enzyme Y216F, pH 9.2 Escherichia coli
6.3.2.4 6.2
-
 D-alanine 37°C, KM-value for D-alanine1 (the first molecule of substrate that is activated by the enzyme molecule), mutant enzyme S150A, pH 6.0 Escherichia coli
6.3.2.4 8
-
 D-alanine 37°C, KM-value for D-alanine2 (the second molecule of substrate that is activated by the enzyme molecule), wild-type enzyme, pH 6.0 Escherichia coli
6.3.2.4 11
-
 D-alanine 37°C, KM-value for D-alanine1 (the first molecule of substrate that is activated by the enzyme molecule), mutant enzyme E15Q, pH 6.0 Escherichia coli
6.3.2.4 16
-
 D-alanine 37°C, KM-value for D-alanine1 (the first molecule of substrate that is activated by the enzyme molecule), mutant enzyme Y216F, pH 7.5 Escherichia coli
6.3.2.4 44
-
 D-alanine 37°C, KM-value for D-alanine1 (the first molecule of substrate that is activated by the enzyme molecule) , mutant enzyme S150A, pH 9.2 Escherichia coli
6.3.2.4 65
-
 D-alanine 37°C, KM-value for D-alanine2 (the second molecule of substrate that is activated by the enzyme molecule), mutant enzyme E15Q, pH 9.2 Escherichia coli
6.3.2.4 74
-
 D-alanine 37°C, KM-value for D-alanine2 (the second molecule of substrate that is activated by the enzyme molecule), mutant enzyme E15Q, pH 7.5 Escherichia coli
6.3.2.4 80
-
 D-alanine 37°C, KM-value for D-alanine2 (the second molecule of substrate that is activated by the enzyme molecule), mutant enzyme S150A, pH 7.5 Escherichia coli
6.3.2.4 82
-
 D-alanine 37°C, KM-value for D-alanine2 (the second molecule of substrate that is activated by the enzyme molecule), mutant enzyme E15Q, pH 6.0 Escherichia coli
6.3.2.4 140
-
 D-alanine 37°C, KM-value for D-alanine2 (the second molecule of substrate that is activated by the enzyme molecule), mutant enzyme S150A, pH 6.0 Escherichia coli
6.3.2.4 150
-
 D-alanine 37°C, KM-value for D-alanine1 (the first molecule of substrate that is activated by the enzyme molecule) , mutant enzyme Y216F, pH 6.0 Escherichia coli
6.3.2.4 450
-
 D-alanine 37°C, KM-value for D-alanine2 (the second molecule of substrate that is activated by the enzyme molecule), mutant enzyme Y216F, pH 6.0 Escherichia coli
6.3.2.4 630
-
 D-alanine 37°C, KM-value for D-alanine2 (the second molecule of substrate that is activated by the enzyme molecule), mutant enzyme Y216F, pH 7.5 Escherichia coli
6.3.2.4 1100
-
 D-alanine 37°C, KM-value for D-alanine2 (the second molecule of substrate that is activated by the enzyme molecule), mutant enzyme S150A, pH 9.2 Escherichia coli
6.3.2.4 1200
-
 D-alanine 37°C, KM-value for D-alanine2 (the second molecule of substrate that is activated by the enzyme molecule), mutant enzyme Y216F, pH 9.2 Escherichia coli

Organism

EC Number Organism UniProt Comment Textmining
6.1.2.1 Escherichia coli
-
-
-
6.3.2.4 Escherichia coli P07862
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
6.1.2.1 D-Ala + D-2-hydroxybutanoate + ATP
-
 Escherichia coli D-Ala-D-2-hydroxybutanoate + ADP + phosphate
-
 ?
6.1.2.1 D-Ala + D-Ala + ATP
-
 Escherichia coli D-Ala-D-Ala + ADP + phosphate
-
 ?
6.1.2.1 D-Ala + D-lactate + ATP
-
 Escherichia coli D-Ala-D-lactate + ADP + phosphate
-
 ?
6.3.2.4 ATP + 2 D-alanine Y216, S150, and E15 form a hydrogen-bonding triad that orients an omega-loop to close over the active site and also to orient substrate D-Ala1 (the first molecule of substrate that is activated by the enzyme protein). The bifunctional enzyme also catalyzes the formation of D-alanyl-D-lactate (D-alanine-(R)-lactate ligase) Escherichia coli ADP + phosphate + D-alanyl-D-alanine
-
 ?

Synonyms

EC Number Synonyms Comment Organism
6.1.2.1 depsipeptide ligase
-
 Escherichia coli
6.3.2.4 D-alanyl-D-alanine ligase B
-
 Escherichia coli
6.3.2.4 DdlB
-
 Escherichia coli

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
6.1.2.1 0.023
-
 D-lactate mutant S150A, pH 6.0, 37°C Escherichia coli
6.1.2.1 0.057
-
 D-lactate mutant S150A, pH 7.5, 37°C Escherichia coli
6.1.2.1 0.13
-
 D-lactate mutant Y216F, pH 6.0, 37°C Escherichia coli
6.1.2.1 0.65
-
 D-lactate isoform VanA, pH 6.0, 37°C Escherichia coli
6.1.2.1 0.67
-
 D-lactate isoform VanA, pH 7.5, 37°C Escherichia coli
6.1.2.1 0.7
-
 D-lactate mutant Y216F, pH 7.5, 37°C Escherichia coli
6.1.2.1 1.17
-
 D-lactate isoform VanA, pH 8.3, 37°C Escherichia coli
6.1.2.1 3.3
-
 D-Ala mutant S150A, pH 6.0, 37°C Escherichia coli
6.1.2.1 7.5
-
 D-Ala mutant Y216F, pH 6.0, 37°C Escherichia coli
6.1.2.1 10.5
-
 D-Ala mutant Y216F, pH 7.5, 37°C Escherichia coli
6.1.2.1 15
-
 D-Ala isoform VanA, pH 8.3, 37°C Escherichia coli
6.1.2.1 15.7
-
 D-Ala isoform VanA, pH 7.5, 37°C Escherichia coli
6.1.2.1 16.3
-
 D-Ala mutant S150A, pH 7.5, 37°C Escherichia coli
6.3.2.4 3.3
-
 D-alanine 37°C, mutant enzyme E15Q, pH 6.0 Escherichia coli
6.3.2.4 3.3
-
 D-alanine 37°C, mutant enzyme S150A, pH 6.0 Escherichia coli
6.3.2.4 7.5
-
 D-alanine 37°C, mutant enzyme Y216F, pH 6.0 Escherichia coli
6.3.2.4 10.5
-
 D-alanine 37°C, mutant enzyme Y216F, pH 7.5 Escherichia coli
6.3.2.4 11.83
-
 D-alanine 37°C, wild-type enzyme, pH 6.0 Escherichia coli
6.3.2.4 13.7
-
 D-alanine 37°C, mutant enzyme E15Q, pH 7.5 Escherichia coli
6.3.2.4 14.7
-
 D-alanine 37°C, wild-type enzyme, pH 7.5 Escherichia coli
6.3.2.4 16.3
-
 D-alanine 37°C, mutant enzyme S150A, pH 7.5 Escherichia coli
6.3.2.4 18.3
-
 D-alanine 37°C, mutant enzyme E15Q, pH 9.2 Escherichia coli
6.3.2.4 18.3
-
 D-alanine 37°C, mutant enzyme S150A, pH 9.2 Escherichia coli
6.3.2.4 20
-
 D-alanine 37°C, mutant enzyme Y216F, pH 9.2 Escherichia coli
6.3.2.4 26.7
-
 D-alanine 37°C, wild-type enzyme, pH 9.2 Escherichia coli

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
6.1.2.1 0.014
-
 D-lactate mutant S150A, pH 6.0, 37°C Escherichia coli
6.1.2.1 0.024
-
 D-Ala mutant S150A, pH 6.0, 37°C Escherichia coli
6.1.2.1 0.025
-
 D-lactate mutant Y216F, pH 6.0, 37°C Escherichia coli
6.1.2.1 0.026
-
 D-lactate mutant S150A, pH 7.5, 37°C Escherichia coli
6.1.2.1 0.026
-
 D-lactate mutant Y216F, pH 7.5, 37°C Escherichia coli
6.1.2.1 0.028
-
 D-Ala isoform VanA, pH 7.5, 37°C Escherichia coli
6.1.2.1 0.05
-
 D-Ala mutant Y216F, pH 6.0, 37°C Escherichia coli
6.1.2.1 0.2
-
 D-Ala mutant S150A, pH 7.5, 37°C Escherichia coli
6.1.2.1 0.21
-
 D-Ala isoform VanA, pH 8.3, 37°C Escherichia coli
6.1.2.1 0.5
-
 D-lactate isoform VanA, pH 6.0, 37°C Escherichia coli
6.1.2.1 0.6
-
 D-lactate isoform VanA, pH 7.5, 37°C Escherichia coli
6.1.2.1 0.65
-
 D-Ala mutant Y216F, pH 7.5, 37°C Escherichia coli
6.1.2.1 0.74
-
 D-lactate isoform VanA, pH 8.3, 37°C Escherichia coli
6.3.2.4 1.4
-
 D-alanine 37°C, mutant enzyme S150A, kcat/Km(D-alanine2), pH 6.0. D-Alanine2 is the second molecule of substrate that is activated by the enzyme molecule Escherichia coli
6.3.2.4 2.4
-
 D-alanine 37°C, mutant enzyme E15Q, kcat/Km(D-alanine2), pH 6.0. D-Alanine2 is the second molecule of substrate that is activated by the enzyme molecule Escherichia coli
6.3.2.4 3
-
 D-alanine 37°C, mutant enzyme Y216F, kcat/Km(D-alanine2), pH 6.0. D-Alanine2 is the second molecule of substrate that is activated by the enzyme molecule Escherichia coli
6.3.2.4 11
-
 D-alanine 37°C, mutant enzyme E15Q, kcat/Km(D-alanine2), pH 7.5. D-Alanine2 is the second molecule of substrate that is activated by the enzyme molecule Escherichia coli
6.3.2.4 12
-
 D-alanine 37°C, mutant enzyme S150A, kcat/Km(D-alanine2), pH 7.5. D-Alanine2 is the second molecule of substrate that is activated by the enzyme molecule Escherichia coli
6.3.2.4 17
-
 D-alanine 37°C, mutant enzyme E15Q, kcat/Km(D-alanine2), pH 9.2. D-Alanine2 is the second molecule of substrate that is activated by the enzyme molecule Escherichia coli
6.3.2.4 26
-
 D-alanine 37°C, mutant enzyme S150A, kcat/Km(D-alanine2), pH 9.2. D-Alanine2 is the second molecule of substrate that is activated by the enzyme molecule Escherichia coli
6.3.2.4 39
-
 D-alanine 37°C, mutant enzyme Y216F, kcat/Km(D-alanine2), pH 7.5. D-Alanine2 is the second molecule of substrate that is activated by the enzyme molecule Escherichia coli
6.3.2.4 89
-
 D-alanine 37°C, wild-type enzyme, kcat/Km(D-alanine2), pH 6.0. D-Alanine2 is the second molecule of substrate that is activated by the enzyme molecule Escherichia coli
6.3.2.4 260
-
 D-alanine 37°C, mutant enzyme Y216F, kcat/Km(D-alanine2), pH 9.2. D-Alanine2 is the second molecule of substrate that is activated by the enzyme molecule Escherichia coli
6.3.2.4 440
-
 D-alanine 37°C, wild-type enzyme, kcat/Km(D-alanine2), pH 7.5. D-Alanine2 is the second molecule of substrate that is activated by the enzyme molecule Escherichia coli
6.3.2.4 1500
-
 D-alanine 37°C, wild-type enzyme, kcat/Km(D-alanine2), pH 9.2. D-Alanine2 is the second molecule of substrate that is activated by the enzyme molecule Escherichia coli