EC Number | Cloned (Comment) | Organism |
---|---|---|
5.1.2.2 | gene mdlA, genetic library construction, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Pseudomonas putida |
EC Number | Protein Variants | Comment | Organism |
---|---|---|---|
5.1.2.2 | additional information | directed evolution of mandelate racemase by a high-throughput screening method, overview | Pseudomonas putida |
5.1.2.2 | V22I | site-directed mutagenesis, kinetic analysis | Pseudomonas putida |
5.1.2.2 | V22I/V29I | site-directed mutagenesis, kinetic analysis | Pseudomonas putida |
5.1.2.2 | V22I/V29I/Y54F | site-directed mutagenesis, the mutant shows 3.5fold greater relative activity as compared to the wild-type enzyme. The enhanced catalytic efficiency mainly arises from the elevated kcat, kinetic analysis | Pseudomonas putida |
5.1.2.2 | V22I/Y54F | site-directed mutagenesis, kinetic analysis | Pseudomonas putida |
5.1.2.2 | V29I | site-directed mutagenesis, kinetic analysis | Pseudomonas putida |
5.1.2.2 | V29I/Y54F | site-directed mutagenesis, kinetic analysis | Pseudomonas putida |
5.1.2.2 | Y54F | site-directed mutagenesis, kinetic analysis | Pseudomonas putida |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
5.1.2.2 | 9.94 | - |
(R)-2-chloromandelate | mutant V29I/Y45F, pH 7.5, 25°C | Pseudomonas putida | |
5.1.2.2 | 10.62 | - |
(S)-2-chloromandelate | mutant V29I/Y45F, pH 7.5, 25°C | Pseudomonas putida | |
5.1.2.2 | 10.9 | - |
(S)-2-chloromandelate | mutant Y45F, pH 7.5, 25°C | Pseudomonas putida | |
5.1.2.2 | 11.66 | - |
(R)-2-chloromandelate | mutant Y45F, pH 7.5, 25°C | Pseudomonas putida | |
5.1.2.2 | 11.99 | - |
(R)-2-chloromandelate | mutant V29I, pH 7.5, 25°C | Pseudomonas putida | |
5.1.2.2 | 13.74 | - |
(S)-2-chloromandelate | mutant V22I/V29I, pH 7.5, 25°C | Pseudomonas putida | |
5.1.2.2 | 14.19 | - |
(S)-2-chloromandelate | mutant V22I/Y45F, pH 7.5, 25°C | Pseudomonas putida | |
5.1.2.2 | 14.35 | - |
(S)-2-chloromandelate | mutant V29I, pH 7.5, 25°C | Pseudomonas putida | |
5.1.2.2 | 14.52 | - |
(R)-2-chloromandelate | mutant V22I/V29I/Y54F, pH 7.5, 25°C | Pseudomonas putida | |
5.1.2.2 | 14.82 | - |
(R)-2-chloromandelate | wild-type enzyme, pH 7.5, 25°C | Pseudomonas putida | |
5.1.2.2 | 15.1 | - |
(R)-2-chloromandelate | mutant V22I/V29I, pH 7.5, 25°C | Pseudomonas putida | |
5.1.2.2 | 15.51 | - |
(R)-2-chloromandelate | mutant V22I, pH 7.5, 25°C | Pseudomonas putida | |
5.1.2.2 | 15.97 | - |
(R)-2-chloromandelate | mutant V22I/Y45F, pH 7.5, 25°C | Pseudomonas putida | |
5.1.2.2 | 17.74 | - |
(S)-2-chloromandelate | wild-type enzyme, pH 7.5, 25°C | Pseudomonas putida | |
5.1.2.2 | 17.94 | - |
(S)-2-chloromandelate | mutant V22I/V29I/Y54F, pH 7.5, 25°C | Pseudomonas putida | |
5.1.2.2 | 19.91 | - |
(S)-2-chloromandelate | mutant V22I, pH 7.5, 25°C | Pseudomonas putida |
EC Number | Metals/Ions | Comment | Organism | Structure |
---|---|---|---|---|
5.1.2.2 | Mg2+ | required | Pseudomonas putida |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
5.1.2.2 | Pseudomonas putida | P11444 | i.e. Arthrobacter siderocapsulatus | - |
5.1.2.2 | Pseudomonas putida ATCC 12633 | P11444 | i.e. Arthrobacter siderocapsulatus | - |
EC Number | Purification (Comment) | Organism |
---|---|---|
5.1.2.2 | recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography | Pseudomonas putida |
EC Number | Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|---|
5.1.2.2 | 0.00057 | - |
wild-type enzyme, pH 7.5, 25°C, substrate (S)-2-chloromandelate | Pseudomonas putida |
5.1.2.2 | 0.00065 | - |
wild-type enzyme, pH 7.5, 25°C, substrate (R)-2-chloromandelate | Pseudomonas putida |
5.1.2.2 | 0.00089 | - |
mutant Y45F, pH 7.5, 25°C, substrate (R)-2-chloromandelate | Pseudomonas putida |
5.1.2.2 | 0.00103 | - |
mutant V29I, pH 7.5, 25°C, substrate (R)-2-chloromandelate | Pseudomonas putida |
5.1.2.2 | 0.00118 | - |
mutant Y45F, pH 7.5, 25°C, substrate (S)-2-chloromandelate | Pseudomonas putida |
5.1.2.2 | 0.00125 | - |
mutant V22I, pH 7.5, 25°C, substrate (R)-2-chloromandelate | Pseudomonas putida |
5.1.2.2 | 0.00133 | - |
mutant V29I, pH 7.5, 25°C, substrate (S)-2-chloromandelate | Pseudomonas putida |
5.1.2.2 | 0.00134 | - |
mutant V29I/Y45F, pH 7.5, 25°C, substrate (S)-2-chloromandelate | Pseudomonas putida |
5.1.2.2 | 0.00143 | - |
mutant V22I, pH 7.5, 25°C, substrate (S)-2-chloromandelate | Pseudomonas putida |
5.1.2.2 | 0.00143 | - |
mutant V22I/Y45F, pH 7.5, 25°C, substrate (R)-2-chloromandelate | Pseudomonas putida |
5.1.2.2 | 0.00149 | - |
mutant V22I/V29I, pH 7.5, 25°C, substrate (S)-2-chloromandelate | Pseudomonas putida |
5.1.2.2 | 0.00152 | - |
mutant V29I/Y45F, pH 7.5, 25°C, substrate (R)-2-chloromandelate | Pseudomonas putida |
5.1.2.2 | 0.00155 | - |
mutant V22I/V29I, pH 7.5, 25°C, substrate (R)-2-chloromandelate | Pseudomonas putida |
5.1.2.2 | 0.00174 | - |
mutant V22I/V29I/Y54F, pH 7.5, 25°C, substrate (R)-2-chloromandelate | Pseudomonas putida |
5.1.2.2 | 0.00175 | - |
mutant V22I/Y45F, pH 7.5, 25°C, substrate (S)-2-chloromandelate | Pseudomonas putida |
5.1.2.2 | 0.00194 | - |
mutant V22I/V29I/Y54F, pH 7.5, 25°C, substrate (S)-2-chloromandelate | Pseudomonas putida |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
5.1.2.2 | (R)-2-chloromandelate | - |
Pseudomonas putida | (S)-2-chloromandelate | - |
r | |
5.1.2.2 | (R)-2-chloromandelate | - |
Pseudomonas putida ATCC 12633 | (S)-2-chloromandelate | - |
r | |
5.1.2.2 | (S)-2-chloromandelate | binding structure | Pseudomonas putida | (R)-2-chloromandelate | - |
r | |
5.1.2.2 | (S)-2-chloromandelate | binding structure | Pseudomonas putida ATCC 12633 | (R)-2-chloromandelate | - |
r | |
5.1.2.2 | additional information | enzyme activity detection in the high-throughput screening method of enzymes involving the racemization of (S)-2-chloromandelate, method evaluation, overview | Pseudomonas putida | ? | - |
? | |
5.1.2.2 | additional information | enzyme activity detection in the high-throughput screening method of enzymes involving the racemization of (S)-2-chloromandelate, method evaluation, overview | Pseudomonas putida ATCC 12633 | ? | - |
? |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
5.1.2.2 | mdlA | - |
Pseudomonas putida |
EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|---|
5.1.2.2 | 25 | - |
assay at | Pseudomonas putida |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
5.1.2.2 | 0.9 | - |
(R)-2-chloromandelate | wild-type enzyme, pH 7.5, 25°C | Pseudomonas putida | |
5.1.2.2 | 0.99 | - |
(S)-2-chloromandelate | wild-type enzyme, pH 7.5, 25°C | Pseudomonas putida | |
5.1.2.2 | 1.23 | - |
(R)-2-chloromandelate | mutant Y45F, pH 7.5, 25°C | Pseudomonas putida | |
5.1.2.2 | 1.33 | - |
(S)-2-chloromandelate | mutant Y45F, pH 7.5, 25°C | Pseudomonas putida | |
5.1.2.2 | 1.45 | - |
(R)-2-chloromandelate | mutant V29I, pH 7.5, 25°C | Pseudomonas putida | |
5.1.2.2 | 1.73 | - |
(R)-2-chloromandelate | mutant V29I/Y45F, pH 7.5, 25°C | Pseudomonas putida | |
5.1.2.2 | 1.82 | - |
(R)-2-chloromandelate | mutant V22I, pH 7.5, 25°C | Pseudomonas putida | |
5.1.2.2 | 1.83 | - |
(S)-2-chloromandelate | mutant V29I, pH 7.5, 25°C | Pseudomonas putida | |
5.1.2.2 | 1.96 | - |
(S)-2-chloromandelate | mutant V29I/Y45F, pH 7.5, 25°C | Pseudomonas putida | |
5.1.2.2 | 2.35 | - |
(S)-2-chloromandelate | mutant V22I/V29I, pH 7.5, 25°C | Pseudomonas putida | |
5.1.2.2 | 2.4 | - |
(R)-2-chloromandelate | mutant V22I/Y45F, pH 7.5, 25°C | Pseudomonas putida | |
5.1.2.2 | 2.47 | - |
(S)-2-chloromandelate | mutant V22I, pH 7.5, 25°C | Pseudomonas putida | |
5.1.2.2 | 2.51 | - |
(S)-2-chloromandelate | mutant V22I/Y45F, pH 7.5, 25°C | Pseudomonas putida | |
5.1.2.2 | 2.57 | - |
(R)-2-chloromandelate | mutant V22I/V29I/Y54F, pH 7.5, 25°C | Pseudomonas putida | |
5.1.2.2 | 2.6 | - |
(R)-2-chloromandelate | mutant V22I/V29I, pH 7.5, 25°C | Pseudomonas putida | |
5.1.2.2 | 3.49 | - |
(S)-2-chloromandelate | mutant V22I/V29I/Y54F, pH 7.5, 25°C | Pseudomonas putida |
EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|---|
5.1.2.2 | 7.5 | - |
assay at | Pseudomonas putida |
EC Number | General Information | Comment | Organism |
---|---|---|---|
5.1.2.2 | additional information | enzyme molecular dynamics simulation and molecular docking | Pseudomonas putida |
EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
5.1.2.2 | 0.056 | - |
(S)-2-chloromandelate | wild-type enzyme, pH 7.5, 25°C | Pseudomonas putida | |
5.1.2.2 | 0.061 | - |
(R)-2-chloromandelate | wild-type enzyme, pH 7.5, 25°C | Pseudomonas putida | |
5.1.2.2 | 0.106 | - |
(R)-2-chloromandelate | mutant Y45F, pH 7.5, 25°C | Pseudomonas putida | |
5.1.2.2 | 0.118 | - |
(R)-2-chloromandelate | mutant V22I, pH 7.5, 25°C | Pseudomonas putida | |
5.1.2.2 | 0.121 | - |
(R)-2-chloromandelate | mutant V29I, pH 7.5, 25°C | Pseudomonas putida | |
5.1.2.2 | 0.122 | - |
(S)-2-chloromandelate | mutant Y45F, pH 7.5, 25°C | Pseudomonas putida | |
5.1.2.2 | 0.124 | - |
(S)-2-chloromandelate | mutant V22I, pH 7.5, 25°C | Pseudomonas putida | |
5.1.2.2 | 0.128 | - |
(S)-2-chloromandelate | mutant V29I, pH 7.5, 25°C | Pseudomonas putida | |
5.1.2.2 | 0.15 | - |
(R)-2-chloromandelate | mutant V22I/Y45F, pH 7.5, 25°C | Pseudomonas putida | |
5.1.2.2 | 0.159 | - |
(S)-2-chloromandelate | mutant V22I/V29I, pH 7.5, 25°C | Pseudomonas putida | |
5.1.2.2 | 0.172 | - |
(R)-2-chloromandelate | mutant V22I/V29I, pH 7.5, 25°C | Pseudomonas putida | |
5.1.2.2 | 0.174 | - |
(R)-2-chloromandelate | mutant V29I/Y45F, pH 7.5, 25°C | Pseudomonas putida | |
5.1.2.2 | 0.177 | - |
(R)-2-chloromandelate | mutant V22I/V29I/Y54F, pH 7.5, 25°C | Pseudomonas putida | |
5.1.2.2 | 0.177 | - |
(S)-2-chloromandelate | mutant V22I/Y45F, pH 7.5, 25°C | Pseudomonas putida | |
5.1.2.2 | 0.185 | - |
(S)-2-chloromandelate | mutant V29I/Y45F, pH 7.5, 25°C | Pseudomonas putida | |
5.1.2.2 | 0.195 | - |
(S)-2-chloromandelate | mutant V22I/V29I/Y54F, pH 7.5, 25°C | Pseudomonas putida |