Cloned (Comment) | Organism |
---|---|
Recombinant MR from is overproduced from Escherichia coli strain BL21(DE3) cells transformed with a pET-15b plasmid containing the MR open reading frame. This construct encodes the MR gene product with an N-terminal hexahistidine tag. Mutants are produced by site-directed mutagenesis | Pseudomonas putida |
Protein Variants | Comment | Organism |
---|---|---|
A25V | Catalytic efficiencies (kcat/Km) for all mutants are reduced between 6- and 40fold with the exception of V22I, V26A, V29L, and V22I/V29L which have near wildtype efficiencies with mandelate | Pseudomonas putida |
A25V | variation of the hydrophobic loop, decrease in catalytic efficiency | Pseudomonas putida |
T24S | Catalytic efficiencies (kcat/Km) for all mutants are reduced between 6- and 40fold with the exception of V22I, V26A, V29L, and V22I/V29L which have near wildtype efficiencies with mandelate | Pseudomonas putida |
T24S | variation of the hydrophobic loop, decrease in catalytic efficiency | Pseudomonas putida |
V22A | Catalytic efficiencies (kcat/Km) for all mutants are reduced between 6- and 40fold with the exception of V22I, V26A, V29L, and V22I/V29L which have near wildtype efficiencies with mandelate | Pseudomonas putida |
V22A | variation of the hydrophobic loop, decrease in catalytic efficiency | Pseudomonas putida |
V22F | Catalytic efficiencies (kcat/Km) for all mutants are reduced between 6- and 40fold with the exception of V22I, V26A, V29L, and V22I/V29L which have near wildtype efficiencies with mandelate | Pseudomonas putida |
V22F | variation of the hydrophobic loop, decrease in catalytic efficiency | Pseudomonas putida |
V22I | Catalytic efficiencies (kcat/Km) for all mutants are reduced between 6- and 40fold with the exception of V22I, V26A, V29L, and V22I/V29L which have near wildtype efficiencies with mandelate | Pseudomonas putida |
V22I | variation of the hydrophobic loop. For mandelate, catalytic efficiency similar to wild-type. For substrate 2-naphthylglycolate, increase in catalytic efficiency | Pseudomonas putida |
V22I/V29L | Catalytic efficiencies (kcat/Km) for all mutants are reduced between 6- and 40fold with the exception of V22I, V26A, V29L, and V22I/V29L which have near wildtype efficiencies with mandelate | Pseudomonas putida |
V22I/V29L | variation of the hydrophobic loop, catalytic efficiency similar to wild-type | Pseudomonas putida |
V26A | Catalytic efficiencies (kcat/Km) for all mutants are reduced between 6- and 40fold with the exception of V22I, V26A, V29L, and V22I/V29L which have near wildtype efficiencies with mandelate | Pseudomonas putida |
V26A | variation of the hydrophobic loop, catalytic efficiency similar to wild-type | Pseudomonas putida |
V26A/V29L | Catalytic efficiencies (kcat/Km) for all mutants are reduced between 6- and 40fold with the exception of V22I, V26A, V29L, and V22I/V29L which have near wildtype efficiencies with mandelate | Pseudomonas putida |
V26A/V29L | variation of the hydrophobic loop, decrease in catalytic efficiency | Pseudomonas putida |
V26F | Catalytic efficiencies (kcat/Km) for all mutants are reduced between 6- and 40fold with the exception of V22I, V26A, V29L, and V22I/V29L which have near wildtype efficiencies with mandelate | Pseudomonas putida |
V26F | variation of the hydrophobic loop, decrease in catalytic efficiency | Pseudomonas putida |
V26L | Catalytic efficiencies (kcat/Km) for all mutants are reduced between 6- and 40fold with the exception of V22I, V26A, V29L, and V22I/V29L which have near wildtype efficiencies with mandelate | Pseudomonas putida |
V26L | variation of the hydrophobic loop, decrease in catalytic efficiency | Pseudomonas putida |
V29A | Catalytic efficiencies (kcat/Km) for all mutants are reduced between 6- and 40fold with the exception of V22I, V26A, V29L, and V22I/V29L which have near wildtype efficiencies with mandelate | Pseudomonas putida |
V29A | variation of the hydrophobic loop, decrease in catalytic efficiency | Pseudomonas putida |
V29F | Catalytic efficiencies (kcat/Km) for all mutants are reduced between 6- and 40fold with the exception of V22I, V26A, V29L, and V22I/V29L which have near wildtype efficiencies with mandelate | Pseudomonas putida |
V29F | variation of the hydrophobic loop, decrease in catalytic efficiency | Pseudomonas putida |
V29L | Catalytic efficiencies (kcat/Km) for all mutants are reduced between 6- and 40fold with the exception of V22I, V26A, V29L, and V22I/V29L which have near wildtype efficiencies with mandelate | Pseudomonas putida |
V29L | variation of the hydrophobic loop. For mandelate, catalytic efficiency similar to wild-type. For substrate 2-naphthylglycolate, increase in catalytic efficiency | Pseudomonas putida |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
2-naphthohydroxamate | competitive inhibition at pH 7.5 | Pseudomonas putida | |
2-naphtholhydroxamate | - |
Pseudomonas putida |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.04 | - |
Mg2+ | pH 7.5, mutant V22A, addition of either 10 mM (R)- or (S)-mandelate | Pseudomonas putida | |
0.04 | - |
Mg2+ | pH 7.5, mutant V22F, addition of either 10 mM (R)- or (S)-mandelate | Pseudomonas putida | |
0.05 | - |
Mg2+ | pH 7.5, mutant V26F, addition of either 10 mM (R)- or (S)-mandelate | Pseudomonas putida | |
0.08 | - |
Mg2+ | pH 7.5, mutant V29A, addition of either 10 mM (R)- or (S)-mandelate | Pseudomonas putida | |
0.12 | - |
Mg2+ | pH 7.5, mutant V26L, addition of either 10 mM (R)- or (S)-mandelate | Pseudomonas putida | |
0.15 | - |
Mg2+ | pH 7.5, mutant V26A, addition of either 10 mM (R)- or (S)-mandelate | Pseudomonas putida | |
0.22 | - |
Mg2+ | pH 7.5, mutant V29F, addition of either 10 mM (R)- or (S)-mandelate | Pseudomonas putida | |
0.29 | - |
Mg2+ | pH 7.5, mutant T24S, addition of either 10 mM (R)- or (S)-mandelate | Pseudomonas putida | |
0.34 | - |
Mg2+ | pH 7.5, wild-type, addition of either 10 mM (R)- or (S)-mandelate | Pseudomonas putida | |
0.47 | - |
Mg2+ | pH 7.5, mutant V29L, addition of either 10 mM (R)- or (S)-mandelate | Pseudomonas putida | |
0.51 | - |
Mg2+ | pH 7.5, mutant V26A/V29L, addition of either 10 mM (R)- or (S)-mandelate | Pseudomonas putida | |
0.55 | - |
(S)-2-naphthylglycolate | mutant V29F, pH 7.5, 25°C | Pseudomonas putida | |
0.55 | - |
(S)-2-naphthylglycolate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V29F | Pseudomonas putida | |
0.6 | - |
(S)-Mandelate | mutant V29L, pH 7.5, 25°C | Pseudomonas putida | |
0.6 | - |
(S)-Mandelate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V29L | Pseudomonas putida | |
0.62 | - |
(S)-2-naphthylglycolate | mutant V22F, pH 7.5, 25°C | Pseudomonas putida | |
0.62 | - |
(S)-2-naphthylglycolate | mutant V26L, pH 7.5, 25°C | Pseudomonas putida | |
0.62 | - |
(S)-2-naphthylglycolate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V22F | Pseudomonas putida | |
0.62 | - |
(S)-2-naphthylglycolate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V26L | Pseudomonas putida | |
0.64 | - |
Mg2+ | pH 7.5, mutant A25V, addition of either 10 mM (R)- or (S)-mandelate | Pseudomonas putida | |
0.74 | - |
(S)-2-naphthylglycolate | mutant V26A, pH 7.5, 25°C | Pseudomonas putida | |
0.74 | - |
(S)-2-naphthylglycolate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V26A | Pseudomonas putida | |
0.8 | - |
(S)-2-naphthylglycolate | mutant V29L, pH 7.5, 25°C | Pseudomonas putida | |
0.8 | - |
(S)-2-naphthylglycolate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V29L | Pseudomonas putida | |
0.9 | - |
(R)-mandelate | mutant V29F, pH 7.5, 25°C | Pseudomonas putida | |
0.9 | - |
(R)-mandelate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V29F | Pseudomonas putida | |
0.91 | - |
(R)-mandelate | mutant V26A, pH 7.5, 25°C | Pseudomonas putida | |
0.91 | - |
(R)-mandelate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V26A | Pseudomonas putida | |
0.93 | - |
(S)-2-naphthylglycolate | mutant V29A, pH 7.5, 25°C | Pseudomonas putida | |
0.93 | - |
(S)-2-naphthylglycolate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V29A | Pseudomonas putida | |
0.97 | - |
(S)-Mandelate | wild-type, pH 7.5, 25°C | Pseudomonas putida | |
0.97 | - |
(S)-Mandelate | pH 7.5 at 25°C, 3.3 mM Mg2+, wild-type | Pseudomonas putida | |
1 | - |
(S)-2-naphthylglycolate | wild-type, pH 7.5, 25°C | Pseudomonas putida | |
1 | - |
(S)-2-naphthylglycolate | pH 7.5 at 25°C, 3.3 mM Mg2+, wild-type | Pseudomonas putida | |
1.07 | - |
(S)-Mandelate | pH 7.5 at 25°C, 20 mM Mg2+, wild-type | Pseudomonas putida | |
1.07 | - |
(S)-Mandelate | wild-type, pH 7.5, 25°C, presence of 20 mM Mg2+ | Pseudomonas putida | |
1.1 | - |
(R)-mandelate | mutant A25V, pH 7.5, 25°C | Pseudomonas putida | |
1.1 | - |
(S)-Mandelate | mutant V26A/V29L, pH 7.5, 25°C | Pseudomonas putida | |
1.1 | - |
(S)-Mandelate | mutant V26F, pH 7.5, 25°C | Pseudomonas putida | |
1.1 | - |
(S)-2-naphthylglycolate | mutant V26F, pH 7.5, 25°C | Pseudomonas putida | |
1.1 | - |
(S)-Mandelate | mutant V26L, pH 7.5, 25°C | Pseudomonas putida | |
1.1 | - |
(S)-Mandelate | mutant V29F, pH 7.5, 25°C | Pseudomonas putida | |
1.1 | - |
(R)-2-naphthylglycolate | mutant V29F, pH 7.5, 25°C | Pseudomonas putida | |
1.1 | - |
(R)-mandelate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant A25V | Pseudomonas putida | |
1.1 | - |
(S)-Mandelate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V26A/V29L | Pseudomonas putida | |
1.1 | - |
(S)-Mandelate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V26F | Pseudomonas putida | |
1.1 | - |
(S)-2-naphthylglycolate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V26F | Pseudomonas putida | |
1.1 | - |
(S)-Mandelate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V26L | Pseudomonas putida | |
1.1 | - |
(S)-Mandelate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V29F | Pseudomonas putida | |
1.1 | - |
(R)-2-naphtylglycolate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V29F | Pseudomonas putida | |
1.2 | - |
(R)-mandelate | wild-type, pH 7.5, 25°C | Pseudomonas putida | |
1.2 | - |
(R)-2-naphthylglycolate | wild-type, pH 7.5, 25°C | Pseudomonas putida | |
1.2 | - |
(R)-mandelate | pH 7.5 at 25°C, 3.3 mM Mg2+, wild-type | Pseudomonas putida | |
1.2 | - |
(R)-2-naphtylglycolate | pH 7.5 at 25°C, 3.3 mM Mg2+, wild-type | Pseudomonas putida | |
1.3 | - |
(R)-2-naphthylglycolate | mutant V26L, pH 7.5, 25°C | Pseudomonas putida | |
1.3 | - |
(R)-2-naphtylglycolate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V26L | Pseudomonas putida | |
1.4 | - |
(S)-2-naphthylglycolate | mutant T24S, pH 7.5, 25°C | Pseudomonas putida | |
1.4 | - |
(R)-mandelate | mutant V26A/V29L, pH 7.5, 25°C | Pseudomonas putida | |
1.4 | - |
(S)-2-naphthylglycolate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant T24S | Pseudomonas putida | |
1.4 | - |
(R)-mandelate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V26A/V29L | Pseudomonas putida | |
1.47 | - |
(R)-mandelate | pH 7.5 at 25°C, 20 mM Mg2+, wild-type | Pseudomonas putida | |
1.47 | - |
(R)-mandelate | wild-type, pH 7.5, 25°C, presence of 20 mM Mg2+ | Pseudomonas putida | |
1.5 | - |
(S)-Mandelate | mutant V22I/V29L, pH 7.5, 25°C, presence of 20 mM Mg2+ | Pseudomonas putida | |
1.5 | - |
(S)-Mandelate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V26A/V29L | Pseudomonas putida | |
1.55 | - |
(S)-2-naphthylglycolate | mutant V22A, pH 7.5, 25°C | Pseudomonas putida | |
1.55 | - |
(S)-2-naphthylglycolate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V22A | Pseudomonas putida | |
1.6 | - |
(R)-2-naphthylglycolate | mutant V29L, pH 7.5, 25°C | Pseudomonas putida | |
1.6 | - |
(S)-2-naphthylglycolate | pH 7.5 at 25°C, 20 mM Mg2+, wild-type | Pseudomonas putida | |
1.6 | - |
(R)-2-naphtylglycolate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V29L | Pseudomonas putida | |
1.6 | - |
(S)-2-naphthylglycolate | wild-type, pH 7.5, 25°C, presence of 20 mM Mg2+ | Pseudomonas putida | |
1.67 | - |
(S)-2-naphthylglycolate | mutant A25V, pH 7.5, 25°C | Pseudomonas putida | |
1.67 | - |
(S)-2-naphthylglycolate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant A25V | Pseudomonas putida | |
1.7 | - |
(S)-Mandelate | mutant V22I, pH 7.5, 25°C, presence of 20 mM Mg2+ | Pseudomonas putida | |
1.7 | - |
(S)-Mandelate | mutant V26A, pH 7.5, 25°C | Pseudomonas putida | |
1.7 | - |
(S)-2-naphthylglycolate | mutant V26A/V29L, pH 7.5, 25°C | Pseudomonas putida | |
1.7 | - |
(R)-2-naphthylglycolate | mutant V26F, pH 7.5, 25°C | Pseudomonas putida | |
1.7 | - |
(R)-mandelate | mutant V26L, pH 7.5, 25°C | Pseudomonas putida | |
1.7 | - |
(R)-mandelate | mutant V29L, pH 7.5, 25°C | Pseudomonas putida | |
1.7 | - |
(S)-Mandelate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V22I | Pseudomonas putida | |
1.7 | - |
(S)-Mandelate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V26A | Pseudomonas putida | |
1.7 | - |
(S)-2-naphthylglycolate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V26A/V29L | Pseudomonas putida | |
1.7 | - |
(R)-2-naphtylglycolate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V26F | Pseudomonas putida | |
1.7 | - |
(R)-mandelate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V26L | Pseudomonas putida | |
1.7 | - |
(R)-mandelate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V29L | Pseudomonas putida | |
1.8 | - |
(S)-Mandelate | mutant A25V, pH 7.5, 25°C | Pseudomonas putida | |
1.8 | - |
(R)-mandelate | mutant V26F, pH 7.5, 25°C | Pseudomonas putida | |
1.8 | - |
(R)-2-naphthylglycolate | mutant V29A, pH 7.5, 25°C | Pseudomonas putida | |
1.8 | - |
(S)-Mandelate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant A25V | Pseudomonas putida | |
1.8 | - |
(R)-mandelate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V26F | Pseudomonas putida | |
1.8 | - |
(R)-2-naphtylglycolate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V29A | Pseudomonas putida | |
1.9 | - |
(R)-2-naphthylglycolate | mutant A25V, pH 7.5, 25°C | Pseudomonas putida | |
1.9 | - |
(R)-2-naphthylglycolate | mutant V22A, pH 7.5, 25°C | Pseudomonas putida | |
1.9 | - |
(R)-2-naphtylglycolate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant A25V | Pseudomonas putida | |
1.9 | - |
(R)-2-naphtylglycolate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V22A | Pseudomonas putida | |
2.1 | - |
(R)-2-naphthylglycolate | mutant V26A, pH 7.5, 25°C | Pseudomonas putida | |
2.1 | - |
(R)-2-naphtylglycolate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V26A | Pseudomonas putida | |
2.2 | - |
(R)-2-naphthylglycolate | mutant V22F, pH 7.5, 25°C | Pseudomonas putida | |
2.2 | - |
(R)-2-naphtylglycolate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V22F | Pseudomonas putida | |
2.4 | - |
(S)-2-naphthylglycolate | mutant V22I/V29L, pH 7.5, 25°C, presence of 20 mM Mg2+ | Pseudomonas putida | |
2.4 | - |
(S)-2-naphthylglycolate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V26A/V29L | Pseudomonas putida | |
2.8 | - |
(R)-mandelate | mutant T24S, pH 7.5, 25°C | Pseudomonas putida | |
2.8 | - |
(R)-mandelate | mutant V22I/V29L, pH 7.5, 25°C, presence of 20 mM Mg2+ | Pseudomonas putida | |
2.8 | - |
(R)-mandelate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant T24S | Pseudomonas putida | |
2.8 | - |
(R)-mandelate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V22I/V29L | Pseudomonas putida | |
2.9 | - |
(R)-mandelate | mutant V22I, pH 7.5, 25°C, presence of 20 mM Mg2+ | Pseudomonas putida | |
2.9 | - |
(S)-2-naphthylglycolate | mutant V22I, pH 7.5, 25°C, presence of 20 mM Mg2+ | Pseudomonas putida | |
2.9 | - |
(R)-mandelate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V22I | Pseudomonas putida | |
2.9 | - |
(S)-2-naphthylglycolate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V22I | Pseudomonas putida | |
3 | - |
(S)-Mandelate | mutant T24S, pH 7.5, 25°C | Pseudomonas putida | |
3 | - |
(S)-Mandelate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant T24S | Pseudomonas putida | |
3.2 | - |
(R)-2-naphthylglycolate | mutant T24S, pH 7.5, 25°C | Pseudomonas putida | |
3.2 | - |
(R)-2-naphtylglycolate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant T24S | Pseudomonas putida | |
3.4 | - |
(S)-Mandelate | mutant V22A, pH 7.5, 25°C | Pseudomonas putida | |
3.4 | - |
(S)-Mandelate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V22A | Pseudomonas putida | |
3.5 | - |
(R)-2-naphtylglycolate | pH 7.5 at 25°C, 20 mM Mg2+, wild-type | Pseudomonas putida | |
3.5 | - |
(R)-2-naphthylglycolate | wild-type, pH 7.5, 25°C, presence of 20 mM Mg2+ | Pseudomonas putida | |
3.7 | - |
(R)-2-naphthylglycolate | mutant V22I/V29L, pH 7.5, 25°C, presence of 20 mM Mg2+ | Pseudomonas putida | |
3.7 | - |
(R)-2-naphtylglycolate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V22I/V29L | Pseudomonas putida | |
3.8 | - |
Mg2+ | pH 7.5, mutant V22I, addition of either 10 mM (R)- or (S)-mandelate | Pseudomonas putida | |
3.9 | - |
(S)-Mandelate | mutant V29A, pH 7.5, 25°C | Pseudomonas putida | |
3.9 | - |
(S)-Mandelate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V29A | Pseudomonas putida | |
4.2 | - |
(R)-2-naphthylglycolate | mutant V22I, pH 7.5, 25°C, presence of 20 mM Mg2+ | Pseudomonas putida | |
4.2 | - |
(R)-2-naphtylglycolate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V22I | Pseudomonas putida | |
4.2 | - |
Mg2+ | pH 7.5, mutant V22I/V29L, addition of either 10 mM (R)- or (S)-mandelate | Pseudomonas putida | |
4.4 | - |
(R)-mandelate | mutant V22A, pH 7.5, 25°C | Pseudomonas putida | |
4.4 | - |
(R)-mandelate | mutant V22F, pH 7.5, 25°C | Pseudomonas putida | |
4.4 | - |
(R)-2-naphthylglycolate | mutant V26A/V29L, pH 7.5, 25°C | Pseudomonas putida | |
4.4 | - |
(R)-mandelate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V22A | Pseudomonas putida | |
4.4 | - |
(R)-mandelate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V22F | Pseudomonas putida | |
4.4 | - |
(R)-2-naphtylglycolate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V26A/V29L | Pseudomonas putida | |
5 | - |
(R)-mandelate | mutant V29A, pH 7.5, 25°C | Pseudomonas putida | |
5 | - |
(R)-mandelate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V29A | Pseudomonas putida | |
6.1 | - |
(S)-Mandelate | mutant V22F, pH 7.5, 25°C | Pseudomonas putida | |
6.1 | - |
(S)-Mandelate | pH 7.5 at 25°C, 3.3 mM Mg2+, mutant V22F | Pseudomonas putida |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | catalysis is Mg2+-dependent | Pseudomonas putida | |
Mg2+ | Km value for wild-type, 0.34 mM | Pseudomonas putida |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
40700 | - |
mutants V22A, V26A and V29A | Pseudomonas putida |
40710 | - |
mutants V26A/V26L and T24S | Pseudomonas putida |
40730 | - |
wild type | Pseudomonas putida |
40740 | - |
mutants V22I, V26L and V29L | Pseudomonas putida |
40760 | - |
mutants A25V and V22I/V29L | Pseudomonas putida |
40780 | - |
mutants V22F, V26F and V29F | Pseudomonas putida |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
(S)-mandelate | Pseudomonas putida | catalyzes the Mg2+-dependent 1,1-proton transfer that interconverts the enantiomers, catalysis is done via a two-base mechanism | (R)-mandelate | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Pseudomonas putida | P11444 | - |
- |
Purification (Comment) | Organism |
---|---|
by metal ion affinity chromatography | Pseudomonas putida |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
(R)-2-naphthylglycolate | - |
Pseudomonas putida | (S)-2-naphthylglycolate | - |
r | |
(R)-2-naphtylglycolate | - |
Pseudomonas putida | (S)-2-naphthylglycolate | - |
r | |
(R)-mandelate | - |
Pseudomonas putida | (S)-mandelate | - |
r | |
(S)-2-naphthylglycolate | - |
Pseudomonas putida | (R)-2-naphthylglycolate | - |
r | |
(S)-2-naphthylglycolate | racemisation | Pseudomonas putida | (R)-2-naphthylglycolate | - |
r | |
(S)-mandelate | - |
Pseudomonas putida | (R)-mandelate | - |
r | |
(S)-mandelate | catalyzes the Mg2+-dependent 1,1-proton transfer that interconverts the enantiomers, catalysis is done via a two-base mechanism | Pseudomonas putida | (R)-mandelate | - |
r |
Synonyms | Comment | Organism |
---|---|---|
mandelate racemase | - |
Pseudomonas putida |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Pseudomonas putida |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2.95 | - |
(S)-2-naphthylglycolate | mutant V22F, pH 7.5, 25°C | Pseudomonas putida | |
2.95 | - |
(S)-2-naphthylglycolate | pH 7.5, 3.3 mM Mg2+, mutant V22F | Pseudomonas putida | |
3 | 6 | (R)-mandelate | mutant V26L, pH 7.5, 25°C | Pseudomonas putida | |
3 | 6 | (R)-mandelate | pH 7.5, 3.3 mM Mg2+, mutant V26L | Pseudomonas putida | |
5.4 | - |
(R)-2-naphthylglycolate | mutant A25V, pH 7.5, 25°C | Pseudomonas putida | |
5.4 | - |
(S)-2-naphthylglycolate | mutant V26F, pH 7.5, 25°C | Pseudomonas putida | |
5.4 | - |
(R)-2-naphtylglycolate | pH 7.5, 3.3 mM Mg2+, mutant A25V | Pseudomonas putida | |
5.4 | - |
(S)-2-naphthylglycolate | pH 7.5, 3.3 mM Mg2+, mutant V26F | Pseudomonas putida | |
5.8 | - |
(S)-2-naphthylglycolate | mutant A25V, pH 7.5, 25°C | Pseudomonas putida | |
5.8 | - |
(S)-2-naphthylglycolate | pH 7.5, 3.3 mM Mg2+, mutant A25V | Pseudomonas putida | |
7.1 | - |
(S)-2-naphthylglycolate | mutant V22I/V29L, pH 7.5, 25°C, presence of 20 mM Mg2+ | Pseudomonas putida | |
7.1 | - |
(S)-2-naphthylglycolate | pH 7.5, 3.3 mM Mg2+, mutant V26A/V29L | Pseudomonas putida | |
7.3 | - |
(R)-2-naphthylglycolate | mutant V26F, pH 7.5, 25°C | Pseudomonas putida | |
7.3 | - |
(R)-2-naphtylglycolate | pH 7.5, 3.3 mM Mg2+, mutant V26F | Pseudomonas putida | |
8.2 | - |
(S)-2-naphthylglycolate | mutant V26L, pH 7.5, 25°C | Pseudomonas putida | |
8.2 | - |
(S)-2-naphthylglycolate | pH 7.5, 3.3 mM Mg2+, mutant V26L | Pseudomonas putida | |
8.9 | - |
(S)-2-naphthylglycolate | mutant T24S, pH 7.5, 25°C | Pseudomonas putida | |
8.9 | - |
(S)-2-naphthylglycolate | pH 7.5, 3.3 mM Mg2+, mutant T24S | Pseudomonas putida | |
9.7 | - |
(S)-2-naphthylglycolate | mutant V29F, pH 7.5, 25°C | Pseudomonas putida | |
9.7 | - |
(S)-2-naphthylglycolate | pH 7.5, 3.3 mM Mg2+, mutant V29F | Pseudomonas putida | |
9.8 | - |
(S)-2-naphthylglycolate | mutant V26A/V29L, pH 7.5, 25°C | Pseudomonas putida | |
9.8 | - |
(S)-2-naphthylglycolate | pH 7.5, 3.3 mM Mg2+, mutant V26A/V29L | Pseudomonas putida | |
10.4 | - |
(R)-2-naphthylglycolate | mutant V22F, pH 7.5, 25°C | Pseudomonas putida | |
10.4 | - |
(R)-2-naphtylglycolate | pH 7.5, 3.3 mM Mg2+, mutant V22F | Pseudomonas putida | |
10.5 | - |
(R)-2-naphthylglycolate | mutant V22I/V29L, pH 7.5, 25°C, presence of 20 mM Mg2+ | Pseudomonas putida | |
10.5 | - |
(R)-2-naphtylglycolate | pH 7.5, 3.3 mM Mg2+, mutant V22I/V29L | Pseudomonas putida | |
11.1 | - |
(S)-2-naphthylglycolate | mutant V22A, pH 7.5, 25°C | Pseudomonas putida | |
11.1 | - |
(S)-2-naphthylglycolate | pH 7.5, 3.3 mM Mg2+, mutant V22A | Pseudomonas putida | |
13 | - |
(R)-mandelate | mutant A25V, pH 7.5, 25°C | Pseudomonas putida | |
13 | - |
(R)-mandelate | pH 7.5, 3.3 mM Mg2+, mutant A25V | Pseudomonas putida | |
14 | - |
(S)-2-naphthylglycolate | mutant V26A, pH 7.5, 25°C | Pseudomonas putida | |
14 | - |
(R)-2-naphthylglycolate | mutant V26L, pH 7.5, 25°C | Pseudomonas putida | |
14 | - |
(S)-2-naphthylglycolate | pH 7.5, 3.3 mM Mg2+, mutant V26A | Pseudomonas putida | |
14 | - |
(R)-2-naphtylglycolate | pH 7.5, 3.3 mM Mg2+, mutant V26L | Pseudomonas putida | |
14.1 | - |
(R)-2-naphthylglycolate | mutant V29F, pH 7.5, 25°C | Pseudomonas putida | |
14.1 | - |
(R)-2-naphtylglycolate | pH 7.5, 3.3 mM Mg2+, mutant V29F | Pseudomonas putida | |
15.6 | - |
(R)-2-naphthylglycolate | mutant V22A, pH 7.5, 25°C | Pseudomonas putida | |
15.6 | - |
(R)-2-naphtylglycolate | pH 7.5, 3.3 mM Mg2+, mutant V22A | Pseudomonas putida | |
18.3 | - |
(R)-2-naphthylglycolate | mutant T24S, pH 7.5, 25°C | Pseudomonas putida | |
18.3 | - |
(R)-2-naphtylglycolate | pH 7.5, 3.3 mM Mg2+, mutant T24S | Pseudomonas putida | |
22 | - |
(S)-Mandelate | mutant A25V, pH 7.5, 25°C | Pseudomonas putida | |
22 | - |
(S)-Mandelate | mutant V26F, pH 7.5, 25°C | Pseudomonas putida | |
22 | - |
(S)-Mandelate | pH 7.5, 3.3 mM Mg2+, mutant A25V | Pseudomonas putida | |
22 | - |
(S)-Mandelate | pH 7.5, 3.3 mM Mg2+, mutant V26F | Pseudomonas putida | |
23 | - |
(R)-2-naphthylglycolate | mutant V26A/V29L, pH 7.5, 25°C | Pseudomonas putida | |
23 | - |
(S)-Mandelate | mutant V26L, pH 7.5, 25°C | Pseudomonas putida | |
23 | - |
(R)-2-naphtylglycolate | pH 7.5, 3.3 mM Mg2+, mutant V26A/V29L | Pseudomonas putida | |
23 | - |
(S)-Mandelate | pH 7.5, 3.3 mM Mg2+, mutant V26L | Pseudomonas putida | |
30 | - |
(R)-2-naphthylglycolate | mutant V26A, pH 7.5, 25°C | Pseudomonas putida | |
30 | - |
(S)-2-naphthylglycolate | mutant V29A, pH 7.5, 25°C | Pseudomonas putida | |
30 | - |
(R)-2-naphtylglycolate | pH 7.5, 3.3 mM Mg2+, mutant V26A | Pseudomonas putida | |
30 | - |
(S)-2-naphthylglycolate | pH 7.5, 3.3 mM Mg2+, mutant V29A | Pseudomonas putida | |
32 | - |
(S)-2-naphthylglycolate | wild-type, pH 7.5, 25°C | Pseudomonas putida | |
32 | - |
(S)-2-naphthylglycolate | pH 7.5, 3.3 mM Mg2+, wild-type | Pseudomonas putida | |
33 | - |
(R)-mandelate | mutant V26F, pH 7.5, 25°C | Pseudomonas putida | |
33 | - |
(R)-mandelate | pH 7.5, 3.3 mM Mg2+, mutant V26F | Pseudomonas putida | |
38.4 | - |
(S)-2-naphthylglycolate | pH 7.5, 20 mM Mg2+, wild-type | Pseudomonas putida | |
38.4 | - |
(S)-2-naphthylglycolate | wild-type, pH 7.5, 25°C, presence of 20 mM Mg2+ | Pseudomonas putida | |
46 | - |
(R)-2-naphthylglycolate | wild-type, pH 7.5, 25°C | Pseudomonas putida | |
46 | - |
(R)-2-naphtylglycolate | pH 7.5, 3.3 mM Mg2+, wild-type | Pseudomonas putida | |
53 | - |
(R)-2-naphthylglycolate | mutant V29A, pH 7.5, 25°C | Pseudomonas putida | |
53 | - |
(S)-Mandelate | mutant V29F, pH 7.5, 25°C | Pseudomonas putida | |
53 | - |
(R)-mandelate | mutant V29F, pH 7.5, 25°C | Pseudomonas putida | |
53 | - |
(R)-2-naphtylglycolate | pH 7.5, 3.3 mM Mg2+, mutant V29A | Pseudomonas putida | |
53 | - |
(S)-Mandelate | pH 7.5, 3.3 mM Mg2+, mutant V29F | Pseudomonas putida | |
53 | - |
(R)-mandelate | pH 7.5, 3.3 mM Mg2+, mutant V29F | Pseudomonas putida | |
65 | - |
(S)-2-naphthylglycolate | mutant V29L, pH 7.5, 25°C | Pseudomonas putida | |
65 | - |
(S)-2-naphthylglycolate | pH 7.5, 3.3 mM Mg2+, mutant V29L | Pseudomonas putida | |
70 | - |
(R)-2-naphtylglycolate | pH 7.5, 20 mM Mg2+, wild-type | Pseudomonas putida | |
70 | - |
(R)-2-naphthylglycolate | wild-type, pH 7.5, 25°C, presence of 20 mM Mg2+ | Pseudomonas putida | |
77 | - |
(S)-Mandelate | mutant V22A, pH 7.5, 25°C | Pseudomonas putida | |
77 | - |
(S)-Mandelate | pH 7.5, 3.3 mM Mg2+ mutant V22A | Pseudomonas putida | |
82 | - |
(R)-mandelate | mutant V22A, pH 7.5, 25°C | Pseudomonas putida | |
82 | - |
(R)-mandelate | pH 7.5, 3.3 mM Mg2+, mutant V22A | Pseudomonas putida | |
89 | - |
(S)-Mandelate | mutant V26A/V29L, pH 7.5, 25°C | Pseudomonas putida | |
89 | - |
(S)-Mandelate | pH 7.5, 3.3 mM Mg2+, mutant V26A/V29L | Pseudomonas putida | |
98 | - |
(S)-Mandelate | mutant V22F, pH 7.5, 25°C | Pseudomonas putida | |
98 | - |
(R)-mandelate | mutant V22F, pH 7.5, 25°C | Pseudomonas putida | |
98 | - |
(S)-Mandelate | pH 7.5, 3.3 mM Mg2+, mutant V22F | Pseudomonas putida | |
98 | - |
(R)-mandelate | pH 7.5, 3.3 mM Mg2+, mutant V22F | Pseudomonas putida | |
101 | - |
(S)-2-naphthylglycolate | mutant V22I, pH 7.5, 25°C, presence of 20 mM Mg2+ | Pseudomonas putida | |
101 | - |
(S)-2-naphthylglycolate | pH 7.5, 3.3 mM Mg2+, mutant V22I | Pseudomonas putida | |
102 | - |
(R)-mandelate | mutant T24S, pH 7.5, 25°C | Pseudomonas putida | |
102 | - |
(R)-mandelate | pH 7.5, 3.3 mM Mg2+, mutant T24S | Pseudomonas putida | |
106 | - |
(R)-mandelate | mutant V26A/V29L, pH 7.5, 25°C | Pseudomonas putida | |
106 | - |
(R)-mandelate | pH 7.5, 3.3 mM Mg2+, mutant V26A/V29L | Pseudomonas putida | |
120 | - |
(S)-Mandelate | mutant T24S, pH 7.5, 25°C | Pseudomonas putida | |
120 | - |
(S)-Mandelate | pH 7.5, 3.3 mM Mg2+, mutant T24S | Pseudomonas putida | |
137 | - |
(R)-2-naphthylglycolate | mutant V29L, pH 7.5, 25°C | Pseudomonas putida | |
137 | - |
(R)-2-naphtylglycolate | pH 7.5, 3.3 mM Mg2+, mutant V29L | Pseudomonas putida | |
147 | - |
(R)-2-naphthylglycolate | mutant V22I, pH 7.5, 25°C, presence of 20 mM Mg2+ | Pseudomonas putida | |
147 | - |
(R)-2-naphtylglycolate | pH 7.5, 3.3 mM Mg2+, mutant V22I | Pseudomonas putida | |
176 | - |
(S)-Mandelate | mutant V29A, pH 7.5, 25°C | Pseudomonas putida | |
176 | - |
(S)-Mandelate | pH 7.5, 3.3 mM Mg2+, mutant V29A | Pseudomonas putida | |
180 | - |
(S)-Mandelate | mutant V29L, pH 7.5, 25°C | Pseudomonas putida | |
180 | - |
(S)-Mandelate | pH 7.5, 3.3 mM Mg2+, mutant V29L | Pseudomonas putida | |
240 | - |
(R)-mandelate | mutant V29A, pH 7.5, 25°C | Pseudomonas putida | |
240 | - |
(R)-mandelate | pH 7.5, 3.3 mM Mg2+, mutant V29A | Pseudomonas putida | |
300 | - |
(R)-mandelate | mutant V29L, pH 7.5, 25°C | Pseudomonas putida | |
300 | - |
(R)-mandelate | pH 7.5, 3.3 mM Mg2+, mutant V29L | Pseudomonas putida | |
304 | - |
(R)-mandelate | mutant V26A, pH 7.5, 25°C | Pseudomonas putida | |
304 | - |
(R)-mandelate | pH 7.5, 3.3 mM Mg2+, mutant V26A | Pseudomonas putida | |
470 | - |
(S)-Mandelate | wild-type, pH 7.5, 25°C | Pseudomonas putida | |
470 | - |
(S)-Mandelate | pH 7.5, 3.3 mM Mg2+, wild-type | Pseudomonas putida | |
492 | - |
(S)-Mandelate | pH 7.5, 20 mM Mg2+, wild-type | Pseudomonas putida | |
492 | - |
(S)-Mandelate | wild-type, pH 7.5, 25°C, presence of 20 mM Mg2+ | Pseudomonas putida | |
550 | - |
(S)-Mandelate | mutant V26A, pH 7.5, 25°C | Pseudomonas putida | |
550 | - |
(S)-Mandelate | pH 7.5, 3.3 mM Mg2+, mutant V26A | Pseudomonas putida | |
552 | - |
(R)-mandelate | wild-type, pH 7.5, 25°C | Pseudomonas putida | |
552 | - |
(R)-mandelate | pH 7.5, 3.3 mM Mg2+, wild-type | Pseudomonas putida | |
620 | - |
(R)-mandelate | pH 7.5, 20 mM Mg2+, wild-type | Pseudomonas putida | |
620 | - |
(R)-mandelate | wild-type, pH 7.5, 25°C, presence of 20 mM Mg2+ | Pseudomonas putida | |
710 | - |
(S)-Mandelate | mutant V22I/V29L, pH 7.5, 25°C, presence of 20 mM Mg2+ | Pseudomonas putida | |
710 | - |
(S)-Mandelate | pH 7.5, 3.3 mM Mg2+, mutant V26A/V29L | Pseudomonas putida | |
740 | - |
(S)-Mandelate | mutant V22I, pH 7.5, 25°C, presence of 20 mM Mg2+ | Pseudomonas putida | |
740 | - |
(S)-Mandelate | pH 7.5, 3.3 mM Mg2+, mutant V22I | Pseudomonas putida | |
1010 | - |
(R)-mandelate | mutant V22I/V29L, pH 7.5, 25°C, presence of 20 mM Mg2+ | Pseudomonas putida | |
1010 | - |
(R)-mandelate | pH 7.5, 3.3 mM Mg2+, mutant V22I/V29L | Pseudomonas putida | |
1150 | - |
(R)-mandelate | mutant V22I, pH 7.5, 25°C, presence of 20 mM Mg2+ | Pseudomonas putida | |
1150 | - |
(R)-mandelate | pH 7.5, 3.3 mM Mg2+, mutant V22I | Pseudomonas putida |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Pseudomonas putida |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.023 | - |
2-naphthohydroxamate | wild-type, 20 mM Mg2+, pH 7.5 | Pseudomonas putida | |
0.023 | - |
2-naphtholhydroxamate | wild-type, pH 7.5, 25°C, presence of 20 mM Mg2+ | Pseudomonas putida | |
0.027 | - |
2-naphtholhydroxamate | wild-type, pH 7.5, 25°C | Pseudomonas putida | |
0.027 | - |
2-naphthohydroxamate | wild-type, 3.3 mM Mg2+, pH 7.5 | Pseudomonas putida | |
0.035 | - |
2-naphthohydroxamate | mutant V29L, 3.3 mM Mg2+, pH 7.5 | Pseudomonas putida | |
0.035 | - |
2-naphtholhydroxamate | mutant V29L, pH 7.5, 25°C | Pseudomonas putida | |
0.112 | - |
2-naphtholhydroxamate | mutant V22I/V29L, pH 7.5, 25°C, presence of 20 mM Mg2+ | Pseudomonas putida | |
0.112 | - |
2-naphthohydroxamate | mutant V29I/V29I, 20 mM Mg2+, pH 7.5 | Pseudomonas putida | |
0.19 | - |
2-naphthohydroxamate | mutant V22I, 3.3 mM Mg2+, pH 7.5 | Pseudomonas putida | |
0.19 | - |
2-naphtholhydroxamate | mutant V22I, pH 7.5, 25°C, presence of 20 mM Mg2+ | Pseudomonas putida |