Literature summary for 6.1.2.1 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.

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Protein Variants

Protein Variants	Comment	Organism
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
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

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism	Structure
0.88	-	D-lactate	isoform VanA, pH 7.5, 37°C	Escherichia coli
1.3	-	D-lactate	isoform VanA, pH 6.0, 37°C	Escherichia coli
1.5	-	D-lactate	isoform VanA, pH 8.3, 37°C	Escherichia coli
1.6	-	D-lactate	mutant S150A, pH 6.0, 37°C	Escherichia coli
2.2	-	D-lactate	mutant S150A, pH 7.5, 37°C	Escherichia coli
5.1	-	D-lactate	mutant Y216F, pH 6.0, 37°C	Escherichia coli
16	-	D-Ala	mutant Y216F, pH 7.5, 37°C	Escherichia coli
27	-	D-lactate	mutant Y216F, pH 7.5, 37°C	Escherichia coli
69	-	D-Ala	isoform VanA, pH 8.3, 37°C	Escherichia coli
80	-	D-Ala	mutant S150A, pH 7.5, 37°C	Escherichia coli
100	-	D-Ala	isoform VanA, above 100 mM, pH 7.5, 37°C	Escherichia coli
140	-	D-Ala	mutant S150A, pH 6.0, 37°C	Escherichia coli
150	-	D-Ala	mutant Y216F, pH 6.0, 37°C	Escherichia coli

Organism

Organism	UniProt	Comment	Textmining
Escherichia coli	-	-	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
D-Ala + D-2-hydroxybutanoate + ATP	-	Escherichia coli	D-Ala-D-2-hydroxybutanoate + ADP + phosphate	-	?
D-Ala + D-Ala + ATP	-	Escherichia coli	D-Ala-D-Ala + ADP + phosphate	-	?
D-Ala + D-lactate + ATP	-	Escherichia coli	D-Ala-D-lactate + ADP + phosphate	-	?

Synonyms

Synonyms	Comment	Organism
depsipeptide ligase	-	Escherichia coli

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism	Structure
0.023	-	D-lactate	mutant S150A, pH 6.0, 37°C	Escherichia coli
0.057	-	D-lactate	mutant S150A, pH 7.5, 37°C	Escherichia coli
0.13	-	D-lactate	mutant Y216F, pH 6.0, 37°C	Escherichia coli
0.65	-	D-lactate	isoform VanA, pH 6.0, 37°C	Escherichia coli
0.67	-	D-lactate	isoform VanA, pH 7.5, 37°C	Escherichia coli
0.7	-	D-lactate	mutant Y216F, pH 7.5, 37°C	Escherichia coli
1.17	-	D-lactate	isoform VanA, pH 8.3, 37°C	Escherichia coli
3.3	-	D-Ala	mutant S150A, pH 6.0, 37°C	Escherichia coli
7.5	-	D-Ala	mutant Y216F, pH 6.0, 37°C	Escherichia coli
10.5	-	D-Ala	mutant Y216F, pH 7.5, 37°C	Escherichia coli
15	-	D-Ala	isoform VanA, pH 8.3, 37°C	Escherichia coli
15.7	-	D-Ala	isoform VanA, pH 7.5, 37°C	Escherichia coli
16.3	-	D-Ala	mutant S150A, pH 7.5, 37°C	Escherichia coli

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism	Structure
0.014	-	D-lactate	mutant S150A, pH 6.0, 37°C	Escherichia coli
0.024	-	D-Ala	mutant S150A, pH 6.0, 37°C	Escherichia coli
0.025	-	D-lactate	mutant Y216F, pH 6.0, 37°C	Escherichia coli
0.026	-	D-lactate	mutant S150A, pH 7.5, 37°C	Escherichia coli
0.026	-	D-lactate	mutant Y216F, pH 7.5, 37°C	Escherichia coli
0.028	-	D-Ala	isoform VanA, pH 7.5, 37°C	Escherichia coli
0.05	-	D-Ala	mutant Y216F, pH 6.0, 37°C	Escherichia coli
0.2	-	D-Ala	mutant S150A, pH 7.5, 37°C	Escherichia coli
0.21	-	D-Ala	isoform VanA, pH 8.3, 37°C	Escherichia coli
0.5	-	D-lactate	isoform VanA, pH 6.0, 37°C	Escherichia coli
0.6	-	D-lactate	isoform VanA, pH 7.5, 37°C	Escherichia coli
0.65	-	D-Ala	mutant Y216F, pH 7.5, 37°C	Escherichia coli
0.74	-	D-lactate	isoform VanA, pH 8.3, 37°C	Escherichia coli

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Release 2023.1 (January 2023)