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Literature summary for 3.4.13.22 extracted from

  • McCafferty, D.G.; Lessard, I.A.; Walsh, C.T.
    Mutational analysis of potential zinc-binding residues in the active site of the enterococcal D-Ala-D-Ala dipeptidase VanX (1997), Biochemistry, 36, 10498-10505.
    View publication on PubMed
Search for more literature for 3.4.13.22

Cloned(Commentary)

Cloned (Comment) Organism
overexpression of wild-type and mutants as maltose-binding-protein fusion proteins Enterococcus sp.

Protein Variants

Protein Variants Comment Organism
C78S/C157S site-directed mutagenesis, reduced activity Enterococcus sp.
D123A site-directed mutagenesis, inactive, nearly no remaining Zn2+ Enterococcus sp.
D145A/E146A site-directed mutagenesis, 6fold lowered kcat compared to wild-type, 11% reduced zinc content compared to wild-type Enterococcus sp.
E181A site-directed mutagenesis, inactive, 21.8% reduced zinc content compared to wild-type Enterococcus sp.
H116A site-directed mutagenesis, inactive, 92.1% reduced zinc content compared to wild-type Enterococcus sp.
H116A/H184A site-directed mutagenesis, inactive, 97.5% reduced zinc content compared to wild-type Enterococcus sp.
H13A site-directed mutagenesis, activity equivalent to wild-type, 8.3% reduced zinc content compared to wild-type Enterococcus sp.
H149A site-directed mutagenesis, equivalent to wild-type Enterococcus sp.
H149A/H150A site-directed mutagenesis, 11fold lowered kcat, 14.9% reduced zinc content compared to wild-type Enterococcus sp.
H150A site-directed mutagenesis, equivalent to wild-type Enterococcus sp.
H184A site-directed mutagenesis, inactive, 99.08% reduced zinc content compared to wild-type Enterococcus sp.

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
1.8
-
 D-Ala-D-Ala pH 8.0, 37°C, recombinant mutant C78S/C157S Enterococcus sp.
2.1
-
 D-Ala-D-Ala pH 8.0, 37°C, recombinant mutant H13A Enterococcus sp.
2.4
-
 D-Ala-D-Ala pH 8.0, 37°C, recombinant mutant H149A/H150A Enterococcus sp.
2.5
-
 D-Ala-D-Ala pH 8.0, 37°C, recombinant wild-type enzyme Enterococcus sp.
3.7
-
 D-Ala-D-Ala pH 8.0, 37°C, recombinant mutant D145A/E146A Enterococcus sp.

Metals/Ions

Metals/Ions Comment Organism Structure
Zn2+ zinc binding residues H184, Asp123 and H116 in the active site, zinc content varies in mutant enzymes Enterococcus sp.

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
D-Ala-D-Ala + H2O Enterococcus sp.
-
 D-Ala + D-Ala
-
 ?

Organism

Organism UniProt Comment Textmining
Enterococcus sp.
-
-
-

Reaction

Reaction Comment Organism Reaction ID
D-Ala-D-Ala + H2O = 2 D-Ala reaction mechanism Enterococcus sp.
a

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
D-Ala-D-Ala + H2O
-
 Enterococcus sp. D-Ala + D-Ala
-
 ?

Synonyms

Synonyms Comment Organism
D-Ala-D-Ala dipeptidase VanX
-
 Enterococcus sp.
M15.011 Merops-ID Enterococcus sp.
VanX
-
 Enterococcus sp.

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at Enterococcus sp.

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.64
-
 D-Ala-D-Ala pH 8.0, 37°C, recombinant mutant C78S/C157S Enterococcus sp.
9.1
-
 D-Ala-D-Ala pH 8.0, 37°C, recombinant mutant H149A/H150A Enterococcus sp.
20.2
-
 D-Ala-D-Ala pH 8.0, 37°C, recombinant mutant D145A/E146A Enterococcus sp.
54.1
-
 D-Ala-D-Ala pH 8.0, 37°C, recombinant mutant C78S/C157S Enterococcus sp.
83
-
 D-Ala-D-Ala pH 8.0, 37°C, recombinant mutant H13A Enterococcus sp.
125
-
 D-Ala-D-Ala pH 8.0, 37°C, recombinant wild-type enzyme Enterococcus sp.

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
 assay at Enterococcus sp.