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Literature summary for 3.5.1.77 extracted from
- Gradually accumulating beneficial mutations to improve the thermostability of N-carbamoyl-D-amino acid amidohydrolase by step-wise evolution (2011), Appl. Microbiol. Biotechnol., 90, 1361-1371.
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| H248Q/T262A | the mutant displays a T50 value of 65°C and a DELTAT50 enhancement of 8°C compared with CDase-M3 (T50 is defined as the temperature at which heat treatment for 15 min reduces the initial activity by 50%) | Ralstonia pickettii |
| Q12L | the mutant displays a T50 value of 63°C and a DELTAT50 enhancement of 6°C compared with CDase-M3 (T50 is defined as the temperature at which heat treatment for 15 min reduces the initial activity by 50%) | Ralstonia pickettii |
| Q12L/Q23L/H248Q/T262A/T263S | the mutant exhibits an increase of 10°C in thermostability compared with the parental enzyme M3 | Ralstonia pickettii |
| Q12L/Q23L/H248Q/T262A/T263S/A273K | the mutant displays a T50 value of 69°C and a DELTAT50 enhancement of 12°C compared with CDase-M3 (T50 is defined as the temperature at which heat treatment for 15 min reduces the initial activity by 50%) | Ralstonia pickettii |
| Q12L/Q23L/H248Q/T262A/T263S/E266D | the mutant displays a T50 value of 68°C and a DELTAT50 enhancement of 11°C compared with CDase-M3 (T50 is defined as the temperature at which heat treatment for 15 min reduces the initial activity by 50%) | Ralstonia pickettii |
| Q12L/Q23L/H248Q/T262A/T263S/M31L | the mutant displays a T50 value of 68°C and a DELTAT50 enhancement of 11°C compared with CDase-M3 (T50 is defined as the temperature at which heat treatment for 15 min reduces the initial activity by 50%) | Ralstonia pickettii |
| Q12L/Q23L/H248Q/T262A/T263S/M31L/N242G | the mutant displays a T50 value of 71°C and a DELTAT50 enhancement of 14°C compared with CDase-M3 (T50 is defined as the temperature at which heat treatment for 15 min reduces the initial activity by 50%) | Ralstonia pickettii |
| Q12L/Q23L/H248Q/T262A/T263S/M31L/Q207E | the mutant displays a T50 value of 71°C and a DELTAT50 enhancement of 14°C compared with CDase-M3 (T50 is defined as the temperature at which heat treatment for 15 min reduces the initial activity by 50%) | Ralstonia pickettii |
| Q12L/Q23L/H248Q/T262A/T263S/N242G | the mutant displays a T50 value of 69°C and a DELTAT50 enhancement of 12°C compared with CDase-M3 (T50 is defined as the temperature at which heat treatment for 15 min reduces the initial activity by 50%) | Ralstonia pickettii |
| Q12L/Q23L/H248Q/T262A/T263S/N242G/A273P | the mutant displays a T50 value of 71°C and a DELTAT50 enhancement of 14°C compared with CDase-M3 (T50 is defined as the temperature at which heat treatment for 15 min reduces the initial activity by 50%) | Ralstonia pickettii |
| Q12L/Q23L/H248Q/T262A/T263S/N242G/T271I/A273P | the mutant displays a T50 value of 72°C and a DELTAT50 enhancement of 15°C compared with CDase-M3 (T50 is defined as the temperature at which heat treatment for 15 min reduces the initial activity by 50%) | Ralstonia pickettii |
| Q12L/Q23L/H248Q/T262A/T263S/N93Y | the mutant displays a T50 value of 68°C and a DELTAT50 enhancement of 11°C compared with CDase-M3 (T50 is defined as the temperature at which heat treatment for 15 min reduces the initial activity by 50%) | Ralstonia pickettii |
| Q12L/Q23L/H248Q/T262A/T263S/Q207E | the mutant displays a T50 value of 68°C and a DELTAT50 enhancement of 11°C compared with CDase-M3 (T50 is defined as the temperature at which heat treatment for 15 min reduces the initial activity by 50%) | Ralstonia pickettii |
| Q12L/Q23L/H248Q/T262A/T263S/Q207E/N242G | the mutant displays a T50 value of 71°C and a DELTAT50 enhancement of 14°C compared with CDase-M3 (T50 is defined as the temperature at which heat treatment for 15 min reduces the initial activity by 50%) | Ralstonia pickettii |
| Q12L/Q23L/H248Q/T262A/T263S/T271I | the mutant displays a T50 value of 69°C and a DELTAT50 enhancement of 12°C compared with CDase-M3 (T50 is defined as the temperature at which heat treatment for 15 min reduces the initial activity by 50%) | Ralstonia pickettii |
| Q12L/Q23L/H248Q/T262A/T263S/T271I/A273K | the mutant displays a T50 value of 71°C and a DELTAT50 enhancement of 14°C compared with CDase-M3 (T50 is defined as the temperature at which heat treatment for 15 min reduces the initial activity by 50%) | Ralstonia pickettii |
| Q12L/Q23L/Q207E/N242G/H248Q/T262A/T263S/E266D/T271I/A273P | the mutant displays a T50 value of 73°C and a DELTAT50 enhancement of 16°C compared with CDase-M3 (T50 is defined as the temperature at which heat treatment for 15 min reduces the initial activity by 50%). The mutant retains over 50% of its initial activity after heat treatment at 57°C for at least 100 min | Ralstonia pickettii |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 1 | - |
N-carbamoyl-D-p-hydroxyphenylglycine | mutant enzyme Q12L/Q23L/Q207E/N242G/H248Q/T262A/T263S/E266D/T271I/A273P, in 100 mM potassium phosphate buffer (pH 7.0), at 40°C | Ralstonia pickettii |  |
| 1.2 | - |
N-carbamoyl-D-p-hydroxyphenylglycine | CDase-M3, in 100 mM potassium phosphate buffer (pH 7.0), at 40°C | Ralstonia pickettii | |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| N-carbamoyl-D-p-hydroxyphenylglycine + H2O | Ralstonia pickettii | - |
D-p-hydroxyphenylglycine + NH3 + CO2 | - |
? | |
| N-carbamoyl-D-p-hydroxyphenylglycine + H2O | Ralstonia pickettii CGMCC1596 | - |
D-p-hydroxyphenylglycine + NH3 + CO2 | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Ralstonia pickettii | - |
- |
- |
| Ralstonia pickettii CGMCC1596 | - |
- |
- |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| N-carbamoyl-D-p-hydroxyphenylglycine + H2O | - |
Ralstonia pickettii | D-p-hydroxyphenylglycine + NH3 + CO2 | - |
? | |
| N-carbamoyl-D-p-hydroxyphenylglycine + H2O | - |
Ralstonia pickettii CGMCC1596 | D-p-hydroxyphenylglycine + NH3 + CO2 | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| DCase-M3 | - |
Ralstonia pickettii |
| N-carbamoyl-D-amino acid amidohydrolase | - |
Ralstonia pickettii |
Temperature Stability [°C]
| Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 57 | - |
the activity of CDase-M3 is almost completely lost after heat treatment for 60 min at 57°C | Ralstonia pickettii |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 6.8 | - |
N-carbamoyl-D-p-hydroxyphenylglycine | mutant enzyme Q12L/Q23L/Q207E/N242G/H248Q/T262A/T263S/E266D/T271I/A273P, in 100 mM potassium phosphate buffer (pH 7.0), at 40°C | Ralstonia pickettii | |
| 7 | - |
N-carbamoyl-D-p-hydroxyphenylglycine | CDase-M3, in 100 mM potassium phosphate buffer (pH 7.0), at 40°C | Ralstonia pickettii | |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 5.8 | - |
N-carbamoyl-D-p-hydroxyphenylglycine | CDase-M3, in 100 mM potassium phosphate buffer (pH 7.0), at 40°C | Ralstonia pickettii | |
| 6.8 | - |
N-carbamoyl-D-p-hydroxyphenylglycine | mutant enzyme Q12L/Q23L/Q207E/N242G/H248Q/T262A/T263S/E266D/T271I/A273P, in 100 mM potassium phosphate buffer (pH 7.0), at 40°C | Ralstonia pickettii | |
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