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Literature summary for 3.1.3.16 extracted from

  • Sacco, E.; Cortes, M.; Josseaume, N.; Rice, L.B.; Mainardi, J.L.; Arthur, M.
    Serine/threonine protein phosphatase-mediated control of the peptidoglycan cross-linking L,D-transpeptidase pathway in Enterococcus faecium (2014), MBio, 5, e01446-14.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.1.3.16

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Enterococcus faecium

Protein Variants

Protein Variants Comment Organism
D136Y the mutant shows decreased catalytic efficiency compared to the wild type enzyme Enterococcus faecium
D18Y inactive Enterococcus faecium
S120R the mutant shows strongly decreased catalytic efficiency compared to the wild type enzyme Enterococcus faecium
T101R the mutant shows severely decreased catalytic efficiency compared to the wild type enzyme Enterococcus faecium

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.66
-
 4-nitrophenyl phosphate wild type enzyme, with 2 mM Mn2+, in 50 mM Tris-HCl (pH 8.0), at 37°C Enterococcus faecium
0.69
-
 4-nitrophenyl phosphate wild type enzyme, with 0.05 mM Mn2+, in 50 mM Tris-HCl (pH 8.0), at 37°C Enterococcus faecium
1.2
-
 4-nitrophenyl phosphate mutant enzyme T101R, with 2 mM Mn2+, in 50 mM Tris-HCl (pH 8.0), at 37°C Enterococcus faecium
1.3
-
 4-nitrophenyl phosphate mutant enzyme S120R, with 2 mM Mn2+, in 50 mM Tris-HCl (pH 8.0), at 37°C Enterococcus faecium
1.5
-
 4-nitrophenyl phosphate mutant enzyme D136Y, with 0.05 mM Mn2+, in 50 mM Tris-HCl (pH 8.0), at 37°C Enterococcus faecium
1.8
-
 4-nitrophenyl phosphate mutant enzyme D136Y, with 2 mM Mn2+, in 50 mM Tris-HCl (pH 8.0), at 37°C Enterococcus faecium
2.6
-
 4-nitrophenyl phosphate mutant enzyme S120R, with 0.05 mM Mn2+, in 50 mM Tris-HCl (pH 8.0), at 37°C Enterococcus faecium

Metals/Ions

Metals/Ions Comment Organism Structure
Mn2+ Mn2+ is required for activity. Reduction of the Mn2+ concentration from 2 mM to 0.05 mM leads to a 100fold decrease in the catalytic efficiency of wild type enzyme Enterococcus faecium

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
phosphorylated penicillin-binding protein + H2O Enterococcus faecium
-
 penicillin-binding protein + phosphate
-
 ?
phosphorylated penicillin-binding protein + H2O Enterococcus faecium D344S
-
 penicillin-binding protein + phosphate
-
 ?
phosphorylated serine/threonine protein kinase + H2O Enterococcus faecium
-
 serine/threonine protein kinase + phosphate
-
 ?
phosphorylated serine/threonine protein kinase + H2O Enterococcus faecium D344S
-
 serine/threonine protein kinase + phosphate
-
 ?

Organism

Organism UniProt Comment Textmining
Enterococcus faecium
-
-
-
Enterococcus faecium D344S
-
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni2+-nitrilotriacetate column chromatography and Superdex 75 gel filtration Enterococcus faecium

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4-nitrophenyl phosphate + H2O
-
 Enterococcus faecium 4-nitrophenol + phosphate
-
 ?
4-nitrophenyl phosphate + H2O
-
 Enterococcus faecium D344S 4-nitrophenol + phosphate
-
 ?
phosphorylated penicillin-binding protein + H2O
-
 Enterococcus faecium penicillin-binding protein + phosphate
-
 ?
phosphorylated penicillin-binding protein + H2O
-
 Enterococcus faecium D344S penicillin-binding protein + phosphate
-
 ?
phosphorylated serine/threonine protein kinase + H2O
-
 Enterococcus faecium serine/threonine protein kinase + phosphate
-
 ?
phosphorylated serine/threonine protein kinase + H2O
-
 Enterococcus faecium D344S serine/threonine protein kinase + phosphate
-
 ?

Synonyms

Synonyms Comment Organism
phosphoprotein phosphatase
-
 Enterococcus faecium
serine/threonine protein phosphatase
-
 Enterococcus faecium
StpA
-
 Enterococcus faecium

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.0015
-
 4-nitrophenyl phosphate mutant enzyme D136Y, with 0.05 mM Mn2+, in 50 mM Tris-HCl (pH 8.0), at 37°C Enterococcus faecium
0.0038
-
 4-nitrophenyl phosphate mutant enzyme S120R, with 0.05 mM Mn2+, in 50 mM Tris-HCl (pH 8.0), at 37°C Enterococcus faecium
0.014
-
 4-nitrophenyl phosphate mutant enzyme T101R, with 2 mM Mn2+, in 50 mM Tris-HCl (pH 8.0), at 37°C Enterococcus faecium
0.2
-
 4-nitrophenyl phosphate wild type enzyme, with 0.05 mM Mn2+, in 50 mM Tris-HCl (pH 8.0), at 37°C Enterococcus faecium
0.47
-
 4-nitrophenyl phosphate mutant enzyme S120R, with 2 mM Mn2+, in 50 mM Tris-HCl (pH 8.0), at 37°C Enterococcus faecium
2.33
-
 4-nitrophenyl phosphate mutant enzyme D136Y, with 2 mM Mn2+, in 50 mM Tris-HCl (pH 8.0), at 37°C Enterococcus faecium
20
-
 4-nitrophenyl phosphate wild type enzyme, with 2 mM Mn2+, in 50 mM Tris-HCl (pH 8.0), at 37°C Enterococcus faecium

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.001
-
 4-nitrophenyl phosphate mutant enzyme D136Y, with 0.05 mM Mn2+, in 50 mM Tris-HCl (pH 8.0), at 37°C Enterococcus faecium
0.0015
-
 4-nitrophenyl phosphate mutant enzyme S120R, with 0.05 mM Mn2+, in 50 mM Tris-HCl (pH 8.0), at 37°C Enterococcus faecium
0.0115
-
 4-nitrophenyl phosphate mutant enzyme T101R, with 2 mM Mn2+, in 50 mM Tris-HCl (pH 8.0), at 37°C Enterococcus faecium
0.28
-
 4-nitrophenyl phosphate wild type enzyme, with 0.05 mM Mn2+, in 50 mM Tris-HCl (pH 8.0), at 37°C Enterococcus faecium
0.37
-
 4-nitrophenyl phosphate mutant enzyme S120R, with 2 mM Mn2+, in 50 mM Tris-HCl (pH 8.0), at 37°C Enterococcus faecium
1.33
-
 4-nitrophenyl phosphate mutant enzyme D136Y, with 2 mM Mn2+, in 50 mM Tris-HCl (pH 8.0), at 37°C Enterococcus faecium
30
-
 4-nitrophenyl phosphate wild type enzyme, with 2 mM Mn2+, in 50 mM Tris-HCl (pH 8.0), at 37°C Enterococcus faecium