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Literature summary extracted from
- Lysine racemase from a lactic acid bacterium, Oenococcus oeni: structural basis of substrate specificity (2012), J. Biochem., 152, 505-508.
Cloned(Commentary)
| EC Number | Cloned (Comment) | Organism |
|---|---|---|
| 5.1.1.1 | gene alr, expression of the enzyme in Escherichia coli strain BL21 | Geobacillus stearothermophilus |
| 5.1.1.5 | gene OEOE0162, DNA and amino acid sequence determination and analysis, expression of the enzyme in Escherichia coli strain BL21 | Oenococcus oeni |
Protein Variants
| EC Number | Protein Variants | Comment | Organism |
|---|---|---|---|
| 5.1.1.1 | I222T | site-directed mutagenesis, the mutant is an alanine racemase with lysine racemization activity | Geobacillus stearothermophilus |
| 5.1.1.1 | I222T/Y354W | site-directed mutagenesis, the double mutant is an alanine racemase with lysine racemization activity | Geobacillus stearothermophilus |
| 5.1.1.1 | Y354W | site-directed mutagenesis, the mutant is an alanine racemase with lysine racemization activity | Geobacillus stearothermophilus |
| 5.1.1.5 | T224I | site-directed mutagenesis, the mutant shows significantly decreased activity compared to the wild-type enzyme | Oenococcus oeni |
| 5.1.1.5 | T224I/W355Y | site-directed mutagenesis, the double mutant shows significantly decreased activity compared to the wild-type enzyme | Oenococcus oeni |
| 5.1.1.5 | W355Y | site-directed mutagenesis, the mutant shows significantly decreased activity compared to the wild-type enzyme | Oenococcus oeni |
KM Value [mM]
| EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 5.1.1.1 | 2.3 | - |
D-alanine | pH 8.0, 30°C, recombinant mutant Y354W | Geobacillus stearothermophilus |  |
| 5.1.1.1 | 2.6 | - |
D-alanine | pH 8.0, 30°C, recombinant mutant I222T | Geobacillus stearothermophilus | |
| 5.1.1.1 | 2.8 | - |
D-alanine | pH 8.0, 30°C, recombinant wild-type enzyme | Geobacillus stearothermophilus | |
| 5.1.1.1 | 4 | - |
D-alanine | pH 8.0, 30°C, recombinant mutant I222T/Y354W | Geobacillus stearothermophilus | |
| 5.1.1.1 | 4.5 | - |
L-alanine | pH 8.0, 30°C, recombinant mutant Y354W | Geobacillus stearothermophilus | |
| 5.1.1.1 | 4.7 | - |
L-alanine | pH 8.0, 30°C, recombinant mutant I222T | Geobacillus stearothermophilus | |
| 5.1.1.1 | 5.1 | - |
L-alanine | pH 8.0, 30°C, recombinant wild-type enzyme | Geobacillus stearothermophilus | |
| 5.1.1.1 | 8.3 | - |
L-alanine | pH 8.0, 30°C, recombinant mutant I222T/Y354W | Geobacillus stearothermophilus | |
| 5.1.1.1 | 18 | - |
D-Lysine | pH 8.0, 30°C, recombinant mutant I222T/Y354W | Geobacillus stearothermophilus | |
| 5.1.1.1 | 32 | - |
L-lysine | pH 8.0, 30°C, recombinant mutant I222T/Y354W | Geobacillus stearothermophilus | |
| 5.1.1.5 | 1 | 2 | D-Lysine | pH 8.0, 30°C, recombinant mutant T224I | Oenococcus oeni | |
| 5.1.1.5 | 2 | 3 | L-lysine | pH 8.0, 30°C, recombinant mutant W355Y | Oenococcus oeni | |
| 5.1.1.5 | 9.8 | - |
D-Lysine | pH 8.0, 30°C, recombinant wild-type enzyme | Oenococcus oeni | |
| 5.1.1.5 | 11 | - |
L-lysine | pH 8.0, 30°C, recombinant wild-type enzyme | Oenococcus oeni | |
| 5.1.1.5 | 15 | - |
L-lysine | pH 8.0, 30°C, recombinant mutant T224I | Oenococcus oeni | |
| 5.1.1.5 | 21 | - |
D-Lysine | pH 8.0, 30°C, recombinant mutant W355Y | Oenococcus oeni | |
| 5.1.1.5 | 22 | - |
D-ornithine | pH 8.0, 30°C, recombinant wild-type enzyme | Oenococcus oeni | |
| 5.1.1.5 | 27 | - |
L-ornithine | pH 8.0, 30°C, recombinant wild-type enzyme | Oenococcus oeni | |
| 5.1.1.5 | 27 | - |
L-lysine | pH 8.0, 30°C, recombinant mutant T224I/W355Y | Oenococcus oeni | |
| 5.1.1.5 | 31 | - |
D-Lysine | pH 8.0, 30°C, recombinant mutant T224I/W355Y | Oenococcus oeni | |
Natural Substrates/ Products (Substrates)
| EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 5.1.1.1 | L-alanine | Geobacillus stearothermophilus | - |
D-alanine | - |
r | |
| 5.1.1.5 | L-lysine | Oenococcus oeni | - |
D-lysine | - |
? | |
| 5.1.1.5 | additional information | Oenococcus oeni | the Oenococcus oeni lysine racemase has a specific activity towards basic amino acids, residues Thr224 and Trp355 are important for enzyme substrate specificity | ? | - |
? | |
Organism
| EC Number | Organism | UniProt | Comment | Textmining |
|---|---|---|---|---|
| 5.1.1.1 | Geobacillus stearothermophilus | P10724 | - |
- |
| 5.1.1.5 | Oenococcus oeni | Q04HB7 | gene OEOE0162 | - |
Substrates and Products (Substrate)
| EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|---|
| 5.1.1.1 | D-alanine | - |
Geobacillus stearothermophilus | L-alanine | - |
r | |
| 5.1.1.1 | D-lysine | enzyme mutant I222T/Y354W, no activity with the wild-type enzyme | Geobacillus stearothermophilus | L-lysine | - |
? | |
| 5.1.1.1 | L-alanine | - |
Geobacillus stearothermophilus | D-alanine | - |
r | |
| 5.1.1.1 | L-lysine | enzyme mutant I222T/Y354W, no activity with the wild-type enzyme | Geobacillus stearothermophilus | D-lysine | - |
? | |
| 5.1.1.1 | additional information | residues Ile222 and Tyr354 are important for the enzyme substrate specificity | Geobacillus stearothermophilus | ? | - |
? | |
| 5.1.1.5 | D-lysine | - |
Oenococcus oeni | L-lysine | - |
r | |
| 5.1.1.5 | D-ornithine | - |
Oenococcus oeni | L-ornithine | - |
r | |
| 5.1.1.5 | L-lysine | - |
Oenococcus oeni | D-lysine | - |
? | |
| 5.1.1.5 | L-lysine | - |
Oenococcus oeni | D-lysine | - |
r | |
| 5.1.1.5 | L-Ornithine | - |
Oenococcus oeni | D-Ornithine | - |
r | |
| 5.1.1.5 | additional information | the Oenococcus oeni lysine racemase has a specific activity towards basic amino acids, residues Thr224 and Trp355 are important for enzyme substrate specificity | Oenococcus oeni | ? | - |
? | |
Temperature Optimum [°C]
| EC Number | Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|---|
| 5.1.1.1 | 30 | - |
assay at | Geobacillus stearothermophilus |
| 5.1.1.5 | 30 | - |
assay at | Oenococcus oeni |
Turnover Number [1/s]
| EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 5.1.1.1 | 2.5 | - |
D-Lysine | pH 8.0, 30°C, recombinant mutant I222T/Y354W | Geobacillus stearothermophilus | |
| 5.1.1.1 | 4.17 | - |
L-lysine | pH 8.0, 30°C, recombinant mutant I222T/Y354W | Geobacillus stearothermophilus | |
| 5.1.1.1 | 517 | - |
D-alanine | pH 8.0, 30°C, recombinant mutant Y354W | Geobacillus stearothermophilus | |
| 5.1.1.1 | 633 | - |
D-alanine | pH 8.0, 30°C, recombinant mutant I222T/Y354W | Geobacillus stearothermophilus | |
| 5.1.1.1 | 983 | - |
D-alanine | pH 8.0, 30°C, recombinant mutant I222T | Geobacillus stearothermophilus | |
| 5.1.1.1 | 1017 | - |
D-alanine | pH 8.0, 30°C, recombinant wild-type enzyme | Geobacillus stearothermophilus | |
| 5.1.1.1 | 1133 | - |
L-alanine | pH 8.0, 30°C, recombinant mutant I222T/Y354W | Geobacillus stearothermophilus | |
| 5.1.1.1 | 1133 | - |
L-alanine | pH 8.0, 30°C, recombinant mutant Y354W | Geobacillus stearothermophilus | |
| 5.1.1.1 | 1417 | - |
L-alanine | pH 8.0, 30°C, recombinant mutant I222T | Geobacillus stearothermophilus | |
| 5.1.1.1 | 1533 | - |
L-alanine | pH 8.0, 30°C, recombinant wild-type enzyme | Geobacillus stearothermophilus | |
| 5.1.1.5 | 0.15 | - |
D-Lysine | pH 8.0, 30°C, recombinant mutant T224I/W355Y | Oenococcus oeni | |
| 5.1.1.5 | 0.16 | - |
L-lysine | pH 8.0, 30°C, recombinant mutant T224I/W355Y | Oenococcus oeni | |
| 5.1.1.5 | 0.28 | - |
D-Lysine | pH 8.0, 30°C, recombinant mutant T224I | Oenococcus oeni | |
| 5.1.1.5 | 0.32 | - |
L-lysine | pH 8.0, 30°C, recombinant mutant T224I | Oenococcus oeni | |
| 5.1.1.5 | 0.88 | - |
D-Lysine | pH 8.0, 30°C, recombinant mutant W355Y | Oenococcus oeni | |
| 5.1.1.5 | 1.03 | - |
L-lysine | pH 8.0, 30°C, recombinant mutant W355Y | Oenococcus oeni | |
| 5.1.1.5 | 1.83 | - |
L-ornithine | pH 8.0, 30°C, recombinant wild-type enzyme | Oenococcus oeni | |
| 5.1.1.5 | 1.83 | - |
D-ornithine | pH 8.0, 30°C, recombinant wild-type enzyme | Oenococcus oeni | |
| 5.1.1.5 | 2.83 | - |
D-Lysine | pH 8.0, 30°C, recombinant wild-type enzyme | Oenococcus oeni | |
| 5.1.1.5 | 3.5 | - |
L-lysine | pH 8.0, 30°C, recombinant wild-type enzyme | Oenococcus oeni | |
pH Optimum
| EC Number | pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|---|
| 5.1.1.1 | 8 | - |
assay at | Geobacillus stearothermophilus |
| 5.1.1.5 | 8 | - |
assay at | Oenococcus oeni |
Cofactor
| EC Number | Cofactor | Comment | Organism | Structure |
|---|---|---|---|---|
| 5.1.1.1 | pyridoxal 5'-phosphate | dependent on | Geobacillus stearothermophilus | |
| 5.1.1.5 | pyridoxal 5'-phosphate | dependent on | Oenococcus oeni | |
kcat/KM [mM/s]
| EC Number | kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|---|
| 5.1.1.1 | 0.13 | - |
L-lysine | pH 8.0, 30°C, recombinant mutant I222T/Y354W | Geobacillus stearothermophilus | |
| 5.1.1.1 | 0.14 | - |
D-Lysine | pH 8.0, 30°C, recombinant mutant I222T/Y354W | Geobacillus stearothermophilus | |
| 5.1.1.1 | 136.7 | - |
L-alanine | pH 8.0, 30°C, recombinant mutant I222T/Y354W | Geobacillus stearothermophilus | |
| 5.1.1.1 | 158.3 | - |
D-alanine | pH 8.0, 30°C, recombinant mutant I222T/Y354W | Geobacillus stearothermophilus | |
| 5.1.1.1 | 216.7 | - |
D-alanine | pH 8.0, 30°C, recombinant mutant Y354W | Geobacillus stearothermophilus | |
| 5.1.1.1 | 250 | - |
L-alanine | pH 8.0, 30°C, recombinant mutant Y354W | Geobacillus stearothermophilus | |
| 5.1.1.1 | 300 | - |
L-alanine | pH 8.0, 30°C, recombinant wild-type enzyme | Geobacillus stearothermophilus | |
| 5.1.1.1 | 300 | - |
L-alanine | pH 8.0, 30°C, recombinant mutant I222T | Geobacillus stearothermophilus | |
| 5.1.1.1 | 366.7 | - |
D-alanine | pH 8.0, 30°C, recombinant wild-type enzyme | Geobacillus stearothermophilus | |
| 5.1.1.1 | 383.3 | - |
D-alanine | pH 8.0, 30°C, recombinant mutant I222T | Geobacillus stearothermophilus | |
| 5.1.1.5 | 0.005 | - |
D-Lysine | pH 8.0, 30°C, recombinant mutant T224I/W355Y | Oenococcus oeni | |
| 5.1.1.5 | 0.006 | - |
L-lysine | pH 8.0, 30°C, recombinant mutant T224I/W355Y | Oenococcus oeni | |
| 5.1.1.5 | 0.023 | - |
D-Lysine | pH 8.0, 30°C, recombinant mutant T224I | Oenococcus oeni | |
| 5.1.1.5 | 0.025 | - |
L-lysine | pH 8.0, 30°C, recombinant mutant T224I | Oenococcus oeni | |
| 5.1.1.5 | 0.042 | - |
D-Lysine | pH 8.0, 30°C, recombinant mutant W355Y | Oenococcus oeni | |
| 5.1.1.5 | 0.045 | - |
L-lysine | pH 8.0, 30°C, recombinant mutant W355Y | Oenococcus oeni | |
| 5.1.1.5 | 0.07 | - |
L-ornithine | pH 8.0, 30°C, recombinant wild-type enzyme | Oenococcus oeni | |
| 5.1.1.5 | 0.08 | - |
D-ornithine | pH 8.0, 30°C, recombinant wild-type enzyme | Oenococcus oeni | |
| 5.1.1.5 | 0.28 | - |
D-Lysine | pH 8.0, 30°C, recombinant wild-type enzyme | Oenococcus oeni | |
| 5.1.1.5 | 0.32 | - |
L-lysine | pH 8.0, 30°C, recombinant wild-type enzyme | Oenococcus oeni | |
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