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Literature summary for 3.5.3.6 extracted from
- Hydrophobic mutagenesis and semi-rational engineering of arginine deiminase for markedly enhanced stability and catalytic efficiency (2015), Appl. Biochem. Biotechnol., 176, 1335-1350 .
Application
| Application | Comment | Organism |
|---|---|---|
| drug development | anti-tumor drug | Pseudomonas plecoglossicida |
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expression in Escherichia coli JM109 | Pseudomonas plecoglossicida |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| D38H/A128T/E296K/H404R/I410L/A276W | compared with mutant enzyme D38H/A128T/E296K/H404R/I410L, the mutant enzyme D38H/A128T/E296K/H404R/I410L/A276W displays tremendous pH and thermal stability | Pseudomonas plecoglossicida |
| D38H/A128T/E296K/H404R/I410L/R243L | compared with mutant enzyme D38H/A128T/E296K/H404R/I410L, the mutant enzyme D38H/A128T/E296K/H404R/I410L/R243L exhibits significantly enhanced specific activity and substrate binding affinity, as illustrated by the dramatically reduced Km value (0.16 mmol/l) and a more favorable free energy of L-arginine binding at physiological conditions | Pseudomonas plecoglossicida |
| D38H/A128T/E296K/H404R/I410L/S245D | compared with mutant enzyme D38H/A128T/E296K/H404R/I410L, mutant enzyme D38H/A128T/E296K/H404R/I410L/S245D shows a significantly increased specific activity and a similar Km value at physiological conditions | Pseudomonas plecoglossicida |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.16 | - |
L-arginine | pH 7.4, 37°C, mutant enzyme A128T/H404/I410L/D38H/E298K/R243L | Pseudomonas plecoglossicida |  |
| 0.52 | - |
L-arginine | pH 7.4, 37°C, mutant enzyme A128T/H404/I410L/D38H/E298K | Pseudomonas plecoglossicida | |
| 0.59 | - |
L-arginine | pH 7.4, 37°C, mutant enzyme A128T/H404/I410L/D38H/E298K/S245D | Pseudomonas plecoglossicida | |
| 0.62 | - |
L-arginine | pH 7.4, 37°C, mutant enzyme A128T/H404/I410L/D38H/E298K/A276W | Pseudomonas plecoglossicida | |
| 0.65 | - |
L-arginine | pH 7.4, 37°C, mutant enzyme A128T/H404/I410L | Pseudomonas plecoglossicida | |
Molecular Weight [Da]
| Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
|---|---|---|---|
| 49000 | - |
SDS-PAGE | Pseudomonas plecoglossicida |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| L-arginine + H2O | Pseudomonas plecoglossicida | - |
L-citrulline + NH3 | - |
? | |
| L-arginine + H2O | Pseudomonas plecoglossicida CGMCC2039 | - |
L-citrulline + NH3 | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Pseudomonas plecoglossicida | A7LKB0 | - |
- |
| Pseudomonas plecoglossicida CGMCC2039 | A7LKB0 | - |
- |
Specific Activity [micromol/min/mg]
| Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
|---|---|---|---|
| 7.2 | - |
pH 7.4, 37°C, mutant enzyme A128T/H404/I410L/D38H/E298K/Q162G | Pseudomonas plecoglossicida |
| 8.34 | - |
pH 7.4, 37°C, mutant enzyme A128T/H404/I410L/D38H/E298K/Q162T | Pseudomonas plecoglossicida |
| 8.53 | - |
pH 7.4, 37°C, mutant enzyme A128T/H404/I410L/D38H/E298K/Q162S | Pseudomonas plecoglossicida |
| 8.75 | - |
pH 7.4, 37°C, mutant enzyme A128T/H404/I410L/D38H/E298K | Pseudomonas plecoglossicida |
| 9.92 | - |
pH 7.4, 37°C, mutant A128T/H404R/I410L | Pseudomonas plecoglossicida |
| 16.75 | - |
pH 7.4, 37°C, mutant D38H/A128T/E296K/H404R/I410L | Pseudomonas plecoglossicida |
| 93.4 | - |
pH 7.4, 37°C, mutant D38H/A128T/E296K/H404R/I410L/A276W | Pseudomonas plecoglossicida |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| L-arginine + H2O | - |
Pseudomonas plecoglossicida | L-citrulline + NH3 | - |
? | |
| L-arginine + H2O | - |
Pseudomonas plecoglossicida CGMCC2039 | L-citrulline + NH3 | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| ? | x * 49000, SDS-PAGE | Pseudomonas plecoglossicida |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| ADI | - |
Pseudomonas plecoglossicida |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 32 | - |
mutant enzyme A128T/H404R/I410L | Pseudomonas plecoglossicida |
| 45 | - |
mutant enzyme D38H/A128T/E296K/H404R/I410L | Pseudomonas plecoglossicida |
| 62 | - |
mutant enzyme D38H/A128T/E296K/H404R/I410L/A276W | Pseudomonas plecoglossicida |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 50 | - |
half life: mutant enzyme A128T/H404R/I410L (3.8 h), mutant enzyme D38H/A128T/E296K/H404R/I410L (6.1 h), mutant enzyme D38H/A128T/E296K/H404R/I410L/A276W (6.6 h) | Pseudomonas plecoglossicida |
| 55 | - |
15 min, 50% loss of activity, mutant enzyme A128T/H404R/I410L | Pseudomonas plecoglossicida |
| 56 | - |
15 min, 50% loss of activity, mutant enzyme D38H/A128T/E296K/H404R/I410L | Pseudomonas plecoglossicida |
| 58 | - |
15 min, 50% loss of activity, mutant enzyme D38H/A128T/E296K/H404R/I410L/A276W | Pseudomonas plecoglossicida |
| 60 | - |
half life: mutant enzyme A128T/H404R/I410L (2.5 min), mutant enzyme D38H/A128T/E296K/H404R/I410L (4 min), mutant enzyme D38H/A128T/E296K/H404R/I410L/A276W (17.5 min) | Pseudomonas plecoglossicida |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 20.94 | - |
L-arginine | pH 7.4, 37°C, mutant enzyme A128T/H404/I410L/D38H/E298K/A276W | Pseudomonas plecoglossicida | |
| 21.27 | - |
L-arginine | pH 7.4, 37°C, mutant enzyme A128T/H404/I410L/D38H/E298K | Pseudomonas plecoglossicida | |
| 22.4 | - |
L-arginine | pH 7.4, 37°C, mutant enzyme A128T/H404/I410L/D38H/E298K/S245D | Pseudomonas plecoglossicida | |
| 27.2 | - |
L-arginine | pH 7.4, 37°C, mutant enzyme A128T/H404/I410L | Pseudomonas plecoglossicida | |
| 30.25 | - |
L-arginine | pH 7.4, 37°C, mutant enzyme A128T/H404/I410L/D38H/E298K/R243L | Pseudomonas plecoglossicida | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 6 | - |
wild-type enzyme | Pseudomonas plecoglossicida |
| 6.5 | - |
mutant enzyme A128T/H404R/I410L, mutant enzyme D38H/A128T/E296K/H404R/I410L, mutant enzyme D38H/A128T/E296K/H404R/I410L/A276W | Pseudomonas plecoglossicida |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 33.77 | - |
L-arginine | pH 7.4, 37°C, mutant enzyme A128T/H404/I410L/D38H/E298K/A276W | Pseudomonas plecoglossicida | |
| 37.97 | - |
L-arginine | pH 7.4, 37°C, mutant enzyme A128T/H404/I410L/D38H/E298K/S245D | Pseudomonas plecoglossicida | |
| 40.93 | - |
L-arginine | pH 7.4, 37°C, mutant enzyme A128T/H404/I410L/D38H/E298K | Pseudomonas plecoglossicida | |
| 41.85 | - |
L-arginine | pH 7.4, 37°C, mutant enzyme A128T/H404/I410L | Pseudomonas plecoglossicida | |
| 189.06 | - |
L-arginine | pH 7.4, 37°C, mutant enzyme A128T/H404/I410L/D38H/E298K/R243L | Pseudomonas plecoglossicida | |
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