Application | Comment | Organism |
---|---|---|
synthesis | the F191W mutant is considered to be useful for the enzymatic production of D-Trp which is an important building block of some therapeutic drugs | Achromobacter xylosoxidans |
Cloned (Comment) | Organism |
---|---|
wild type and mutant enzymes are expressed in Escherichia coli JM109 cells | Achromobacter xylosoxidans |
Protein Variants | Comment | Organism |
---|---|---|
F191W | the catalytic efficiency of the mutant toward N-acetyl-D-Trp and N-acetyl-D-Ala is enhanced by 15.6 and 1.5folds, respectively, compared with that of the wild type enzyme. The mutant retains its catalytic efficiency toward N-acetyl-D-Met compared with that of the wild type enzyme | Achromobacter xylosoxidans |
L298A | the catalytic efficiency of the mutant toward N-acetyl-D-Trp is enhanced by 4.4folds compared with that of the wild type enzyme. The catalytic efficiency for N-acetyl-D-Met and N-acetyl-D-Ala decrease to between 2.5 and 5% of that of the wild type enzyme, respectively | Achromobacter xylosoxidans |
L298F | the mutant shows rather lower activities toward all substrates tested including N-acetyl-D-Met compared with the wild type enzyme | Achromobacter xylosoxidans |
M346F | the mutant shows rather lower activities toward all substrates tested including N-acetyl-D-Met compared with the wild type enzyme | Achromobacter xylosoxidans |
V297F | the mutant shows partially lower activities toward all substrates tested compared with the wild type enzyme | Achromobacter xylosoxidans |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
2.03 | - |
N-Acetyl-D-Met | mutant enzyme F191W, in 100 mM HEPES (pH 7.0), at 30°C | Achromobacter xylosoxidans | |
2.97 | - |
N-Acetyl-D-Trp | mutant enzyme F191W, in 100 mM HEPES (pH 7.0), at 30°C | Achromobacter xylosoxidans | |
4.8 | - |
N-Acetyl-D-Met | wild type enzyme, in 100 mM HEPES (pH 7.0), at 30°C | Achromobacter xylosoxidans | |
6.23 | - |
N-Acetyl-D-Trp | wild type enzyme, in 100 mM HEPES (pH 7.0), at 30°C | Achromobacter xylosoxidans | |
6.36 | - |
N-Acetyl-D-Trp | mutant enzyme L298A, in 100 mM HEPES (pH 7.0), at 30°C | Achromobacter xylosoxidans | |
38.61 | - |
N-Acetyl-D-Ala | mutant enzyme F191W, in 100 mM HEPES (pH 7.0), at 30°C | Achromobacter xylosoxidans | |
41 | - |
N-Acetyl-D-Ala | wild type enzyme, in 100 mM HEPES (pH 7.0), at 30°C | Achromobacter xylosoxidans | |
56.4 | - |
N-Acetyl-D-Met | mutant enzyme L298A, in 100 mM HEPES (pH 7.0), at 30°C | Achromobacter xylosoxidans | |
330 | - |
N-Acetyl-D-Ala | mutant enzyme L298A, in 100 mM HEPES (pH 7.0), at 30°C | Achromobacter xylosoxidans |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
52000 | - |
x * 52000, SDS-PAGE | Achromobacter xylosoxidans |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Achromobacter xylosoxidans | - |
subsp. xylosoxydans | - |
Achromobacter xylosoxidans A-6 | - |
subsp. xylosoxydans | - |
Purification (Comment) | Organism |
---|---|
DEAE-Toyopearl column chromatography and butyl-Cellulofine column chromatography | Achromobacter xylosoxidans |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
N-Acetyl-D-Ala + H2O | - |
Achromobacter xylosoxidans | acetate + D-Ala | - |
? | |
N-Acetyl-D-Ala + H2O | - |
Achromobacter xylosoxidans A-6 | acetate + D-Ala | - |
? | |
N-Acetyl-D-Met + H2O | preferred substrate | Achromobacter xylosoxidans | acetate + D-Met | - |
? | |
N-Acetyl-D-Met + H2O | preferred substrate | Achromobacter xylosoxidans A-6 | acetate + D-Met | - |
? | |
N-Acetyl-D-Trp + H2O | poor substrate | Achromobacter xylosoxidans | acetate + D-Trp | - |
? | |
N-Acetyl-D-Trp + H2O | poor substrate | Achromobacter xylosoxidans A-6 | acetate + D-Trp | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 52000, SDS-PAGE | Achromobacter xylosoxidans |
Synonyms | Comment | Organism |
---|---|---|
D-Aminoacylase | - |
Achromobacter xylosoxidans |
D-NAase | - |
Achromobacter xylosoxidans |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
5.97 | - |
N-Acetyl-D-Ala | mutant enzyme L298A, in 100 mM HEPES (pH 7.0), at 30°C | Achromobacter xylosoxidans | |
89.5 | - |
N-Acetyl-D-Trp | wild type enzyme, in 100 mM HEPES (pH 7.0), at 30°C | Achromobacter xylosoxidans | |
229 | - |
N-Acetyl-D-Ala | wild type enzyme, in 100 mM HEPES (pH 7.0), at 30°C | Achromobacter xylosoxidans | |
330 | - |
N-Acetyl-D-Ala | mutant enzyme F191W, in 100 mM HEPES (pH 7.0), at 30°C | Achromobacter xylosoxidans | |
404 | - |
N-Acetyl-D-Trp | mutant enzyme L298A, in 100 mM HEPES (pH 7.0), at 30°C | Achromobacter xylosoxidans | |
463 | - |
N-Acetyl-D-Met | mutant enzyme L298A, in 100 mM HEPES (pH 7.0), at 30°C | Achromobacter xylosoxidans | |
669 | - |
N-Acetyl-D-Trp | mutant enzyme F191W, in 100 mM HEPES (pH 7.0), at 30°C | Achromobacter xylosoxidans | |
829 | - |
N-Acetyl-D-Met | mutant enzyme F191W, in 100 mM HEPES (pH 7.0), at 30°C | Achromobacter xylosoxidans | |
1826 | - |
N-Acetyl-D-Met | wild type enzyme, in 100 mM HEPES (pH 7.0), at 30°C | Achromobacter xylosoxidans |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.02 | - |
N-Acetyl-D-Ala | mutant enzyme L298A, in 100 mM HEPES (pH 7.0), at 30°C | Achromobacter xylosoxidans | |
5.58 | - |
N-Acetyl-D-Ala | wild type enzyme, in 100 mM HEPES (pH 7.0), at 30°C | Achromobacter xylosoxidans | |
8.21 | - |
N-Acetyl-D-Met | mutant enzyme L298A, in 100 mM HEPES (pH 7.0), at 30°C | Achromobacter xylosoxidans | |
8.56 | - |
N-Acetyl-D-Ala | mutant enzyme F191W, in 100 mM HEPES (pH 7.0), at 30°C | Achromobacter xylosoxidans | |
14.4 | - |
N-Acetyl-D-Trp | wild type enzyme, in 100 mM HEPES (pH 7.0), at 30°C | Achromobacter xylosoxidans | |
63.5 | - |
N-Acetyl-D-Trp | mutant enzyme L298A, in 100 mM HEPES (pH 7.0), at 30°C | Achromobacter xylosoxidans | |
225 | - |
N-Acetyl-D-Trp | mutant enzyme F191W, in 100 mM HEPES (pH 7.0), at 30°C | Achromobacter xylosoxidans | |
381 | - |
N-Acetyl-D-Met | wild type enzyme, in 100 mM HEPES (pH 7.0), at 30°C | Achromobacter xylosoxidans | |
407 | - |
N-Acetyl-D-Met | mutant enzyme F191W, in 100 mM HEPES (pH 7.0), at 30°C | Achromobacter xylosoxidans |