Cloned (Comment) | Organism |
---|---|
gene alr, expression of the enzyme in Escherichia coli strain BL21 | Geobacillus stearothermophilus |
Protein Variants | Comment | Organism |
---|---|---|
I222T | site-directed mutagenesis, the mutant is an alanine racemase with lysine racemization activity | Geobacillus stearothermophilus |
I222T/Y354W | site-directed mutagenesis, the double mutant is an alanine racemase with lysine racemization activity | Geobacillus stearothermophilus |
Y354W | site-directed mutagenesis, the mutant is an alanine racemase with lysine racemization activity | Geobacillus stearothermophilus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2.3 | - |
D-alanine | pH 8.0, 30°C, recombinant mutant Y354W | Geobacillus stearothermophilus | |
2.6 | - |
D-alanine | pH 8.0, 30°C, recombinant mutant I222T | Geobacillus stearothermophilus | |
2.8 | - |
D-alanine | pH 8.0, 30°C, recombinant wild-type enzyme | Geobacillus stearothermophilus | |
4 | - |
D-alanine | pH 8.0, 30°C, recombinant mutant I222T/Y354W | Geobacillus stearothermophilus | |
4.5 | - |
L-alanine | pH 8.0, 30°C, recombinant mutant Y354W | Geobacillus stearothermophilus | |
4.7 | - |
L-alanine | pH 8.0, 30°C, recombinant mutant I222T | Geobacillus stearothermophilus | |
5.1 | - |
L-alanine | pH 8.0, 30°C, recombinant wild-type enzyme | Geobacillus stearothermophilus | |
8.3 | - |
L-alanine | pH 8.0, 30°C, recombinant mutant I222T/Y354W | Geobacillus stearothermophilus | |
18 | - |
D-Lysine | pH 8.0, 30°C, recombinant mutant I222T/Y354W | Geobacillus stearothermophilus | |
32 | - |
L-lysine | pH 8.0, 30°C, recombinant mutant I222T/Y354W | Geobacillus stearothermophilus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-alanine | Geobacillus stearothermophilus | - |
D-alanine | - |
r |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Geobacillus stearothermophilus | P10724 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-alanine | - |
Geobacillus stearothermophilus | L-alanine | - |
r | |
D-lysine | enzyme mutant I222T/Y354W, no activity with the wild-type enzyme | Geobacillus stearothermophilus | L-lysine | - |
? | |
L-alanine | - |
Geobacillus stearothermophilus | D-alanine | - |
r | |
L-lysine | enzyme mutant I222T/Y354W, no activity with the wild-type enzyme | Geobacillus stearothermophilus | D-lysine | - |
? | |
additional information | residues Ile222 and Tyr354 are important for the enzyme substrate specificity | Geobacillus stearothermophilus | ? | - |
? |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Geobacillus stearothermophilus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2.5 | - |
D-Lysine | pH 8.0, 30°C, recombinant mutant I222T/Y354W | Geobacillus stearothermophilus | |
4.17 | - |
L-lysine | pH 8.0, 30°C, recombinant mutant I222T/Y354W | Geobacillus stearothermophilus | |
517 | - |
D-alanine | pH 8.0, 30°C, recombinant mutant Y354W | Geobacillus stearothermophilus | |
633 | - |
D-alanine | pH 8.0, 30°C, recombinant mutant I222T/Y354W | Geobacillus stearothermophilus | |
983 | - |
D-alanine | pH 8.0, 30°C, recombinant mutant I222T | Geobacillus stearothermophilus | |
1017 | - |
D-alanine | pH 8.0, 30°C, recombinant wild-type enzyme | Geobacillus stearothermophilus | |
1133 | - |
L-alanine | pH 8.0, 30°C, recombinant mutant I222T/Y354W | Geobacillus stearothermophilus | |
1133 | - |
L-alanine | pH 8.0, 30°C, recombinant mutant Y354W | Geobacillus stearothermophilus | |
1417 | - |
L-alanine | pH 8.0, 30°C, recombinant mutant I222T | Geobacillus stearothermophilus | |
1533 | - |
L-alanine | pH 8.0, 30°C, recombinant wild-type enzyme | Geobacillus stearothermophilus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Geobacillus stearothermophilus |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
pyridoxal 5'-phosphate | dependent on | Geobacillus stearothermophilus |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.13 | - |
L-lysine | pH 8.0, 30°C, recombinant mutant I222T/Y354W | Geobacillus stearothermophilus | |
0.14 | - |
D-Lysine | pH 8.0, 30°C, recombinant mutant I222T/Y354W | Geobacillus stearothermophilus | |
136.7 | - |
L-alanine | pH 8.0, 30°C, recombinant mutant I222T/Y354W | Geobacillus stearothermophilus | |
158.3 | - |
D-alanine | pH 8.0, 30°C, recombinant mutant I222T/Y354W | Geobacillus stearothermophilus | |
216.7 | - |
D-alanine | pH 8.0, 30°C, recombinant mutant Y354W | Geobacillus stearothermophilus | |
250 | - |
L-alanine | pH 8.0, 30°C, recombinant mutant Y354W | Geobacillus stearothermophilus | |
300 | - |
L-alanine | pH 8.0, 30°C, recombinant wild-type enzyme | Geobacillus stearothermophilus | |
300 | - |
L-alanine | pH 8.0, 30°C, recombinant mutant I222T | Geobacillus stearothermophilus | |
366.7 | - |
D-alanine | pH 8.0, 30°C, recombinant wild-type enzyme | Geobacillus stearothermophilus | |
383.3 | - |
D-alanine | pH 8.0, 30°C, recombinant mutant I222T | Geobacillus stearothermophilus |