Protein Variants | Comment | Organism |
---|---|---|
F488G/R526V/T560W | 1.55fold increase in activity with 4-nitrophenyl dodecanoate compared to activity of mutant R526V/T560W | Aeropyrum pernix |
additional information | the esterase activity of the mutant R526V (this mutation transforms a promiscuous acylaminoacyl peptidase into a specific carboxylesterase) towards substrates with long acyl chains is enhanced by protein engineering and solvent optimization. The substrate preference of the enzyme can be further changed from 4-nitrophenyl octanoate to 4-nitrophenyl dodecanoate by protein and solvent engineering | Aeropyrum pernix |
R526V | mutant enzyme with high esterase activity, extreme thermal stability, and high tolerance to organic solvents | Aeropyrum pernix |
R526V/T560W | 1.5fold increase in activity with 4-nitrophenyl dodecanoate compared to activity of mutant R526V | Aeropyrum pernix |
W474V/F488G/R526V/T560W | the mutant enzyme has 7fold higher catalytic efficiency (kcat/Km) for 4-nitrophenyl dodecanoate than the mutant enzyme R526V | Aeropyrum pernix |
W474V/R526V/T560W | 3.11fold increase in activity with 4-nitrophenyl dodecanoate compared to activity of mutant R526V/T560W | Aeropyrum pernix |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00075 | - |
4-nitrophenyl dodecanoate | pH 8.0, 60°C, mutant enzyme W474Q/R526V/T560W | Aeropyrum pernix | |
0.00076 | - |
4-nitrophenyl dodecanoate | pH 8.0, 60°C, mutant enzyme F488Y/R526V/T560W | Aeropyrum pernix | |
0.00101 | - |
4-nitrophenyl dodecanoate | pH 8.0, 60°C, mutant enzyme F488W/R526V/T560W | Aeropyrum pernix | |
0.00122 | - |
4-nitrophenyl dodecanoate | pH 8.0, 60°C, mutant enzyme R526V/T560W | Aeropyrum pernix | |
0.00183 | - |
4-nitrophenyl dodecanoate | pH 8.0, 60°C, mutant enzyme W474A/R526V/T560W | Aeropyrum pernix | |
0.00234 | - |
4-nitrophenyl dodecanoate | pH 8.0, 60°C, mutant enzyme W474V/F488G/R526V/T560W | Aeropyrum pernix | |
0.00243 | - |
4-nitrophenyl dodecanoate | pH 8.0, 60°C, mutant enzyme W474V/R526V/T560W | Aeropyrum pernix | |
0.00388 | - |
4-nitrophenyl dodecanoate | pH 8.0, 60°C, mutant enzyme F488P/R526V/T560W | Aeropyrum pernix | |
0.00579 | - |
4-nitrophenyl dodecanoate | pH 8.0, 60°C, mutant enzyme F488S/R526V/T560W | Aeropyrum pernix | |
0.00737 | - |
4-nitrophenyl dodecanoate | pH 8.0, 60°C, mutant enzyme F488A/R526V/T560W | Aeropyrum pernix | |
0.00834 | - |
4-nitrophenyl dodecanoate | pH 8.0, 60°C, mutant enzyme F488G/R526V/T560W | Aeropyrum pernix | |
0.00862 | - |
4-nitrophenyl dodecanoate | pH 8.0, 60°C, wild-type enzyme | Aeropyrum pernix | |
0.0118 | - |
4-nitrophenyl dodecanoate | pH 8.0, 60°C, mutant enzyme R526V | Aeropyrum pernix |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Aeropyrum pernix | Q9YBQ2 | - |
- |
Aeropyrum pernix DSM 11879 | Q9YBQ2 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-nitrophenyl dodecanoate + H2O | switch of substrate specificity of hyperthermophilic promiscuous acylaminoacyl peptidase by combination of protein and solvent engineering into a specific carboxylesterase | Aeropyrum pernix | 4-nitrophenol + dodecanoate | - |
? | |
4-nitrophenyl dodecanoate + H2O | switch of substrate specificity of hyperthermophilic promiscuous acylaminoacyl peptidase by combination of protein and solvent engineering into a specific carboxylesterase | Aeropyrum pernix DSM 11879 | 4-nitrophenol + dodecanoate | - |
? |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.35 | - |
4-nitrophenyl dodecanoate | pH 8.0, 60°C, wild-type enzyme | Aeropyrum pernix | |
1.73 | - |
4-nitrophenyl dodecanoate | pH 8.0, 60°C, mutant enzyme F488W/R526V/T560W | Aeropyrum pernix | |
2.05 | - |
4-nitrophenyl dodecanoate | pH 8.0, 60°C, mutant enzyme F488Y/R526V/T560W | Aeropyrum pernix | |
5.29 | - |
4-nitrophenyl dodecanoate | pH 8.0, 60°C, mutant enzyme W474Q/R526V/T560W | Aeropyrum pernix | |
5.47 | - |
4-nitrophenyl dodecanoate | pH 8.0, 60°C, mutant enzyme R526V | Aeropyrum pernix | |
7.96 | - |
4-nitrophenyl dodecanoate | pH 8.0, 60°C, mutant enzyme R526V/T560W | Aeropyrum pernix | |
8.17 | - |
4-nitrophenyl dodecanoate | pH 8.0, 60°C, mutant enzyme F488P/R526V/T560W | Aeropyrum pernix | |
10.45 | - |
4-nitrophenyl dodecanoate | pH 8.0, 60°C, mutant enzyme F488S/R526V/T560W | Aeropyrum pernix | |
12.08 | - |
4-nitrophenyl dodecanoate | pH 8.0, 60°C, mutant enzyme F488A/R526V/T560W | Aeropyrum pernix | |
12.61 | - |
4-nitrophenyl dodecanoate | pH 8.0, 60°C, mutant enzyme W474A/R526V/T560W | Aeropyrum pernix | |
17.17 | - |
4-nitrophenyl dodecanoate | pH 8.0, 60°C, mutant enzyme F488G/R526V/T560W | Aeropyrum pernix | |
23.5 | - |
4-nitrophenyl dodecanoate | pH 8.0, 60°C, mutant enzyme W474V/R526V/T560W | Aeropyrum pernix | |
26.25 | - |
4-nitrophenyl dodecanoate | pH 8.0, 60°C, mutant enzyme W474V/F488G/R526V/T560W | Aeropyrum pernix |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
157.2 | - |
4-nitrophenyl dodecanoate | pH 8.0, 60°C, wild-type enzyme | Aeropyrum pernix | |
462.5 | - |
4-nitrophenyl dodecanoate | pH 8.0, 60°C, mutant enzyme R526V | Aeropyrum pernix | |
1638 | - |
4-nitrophenyl dodecanoate | pH 8.0, 60°C, mutant enzyme F488A/R526V/T560W | Aeropyrum pernix | |
1712 | - |
4-nitrophenyl dodecanoate | pH 8.0, 60°C, mutant enzyme F488W/R526V/T560W | Aeropyrum pernix | |
1804 | - |
4-nitrophenyl dodecanoate | pH 8.0, 60°C, mutant enzyme F488S/R526V/T560W | Aeropyrum pernix | |
2059 | - |
4-nitrophenyl dodecanoate | pH 8.0, 60°C, mutant enzyme F488G/R526V/T560W | Aeropyrum pernix | |
2107 | - |
4-nitrophenyl dodecanoate | pH 8.0, 60°C, mutant enzyme F488P/R526V/T560W | Aeropyrum pernix | |
2703 | - |
4-nitrophenyl dodecanoate | pH 8.0, 60°C, mutant enzyme F488Y/R526V/T560W | Aeropyrum pernix | |
6528 | - |
4-nitrophenyl dodecanoate | pH 8.0, 60°C, mutant enzyme R526V/T560W | Aeropyrum pernix | |
6876 | - |
4-nitrophenyl dodecanoate | pH 8.0, 60°C, mutant enzyme W474A/R526V/T560W | Aeropyrum pernix | |
7018 | - |
4-nitrophenyl dodecanoate | pH 8.0, 60°C, mutant enzyme W474Q/R526V/T560W | Aeropyrum pernix | |
9662 | - |
4-nitrophenyl dodecanoate | pH 8.0, 60°C, mutant enzyme W474V/R526V/T560W | Aeropyrum pernix | |
11220 | - |
4-nitrophenyl dodecanoate | pH 8.0, 60°C, mutant enzyme W474V/F488G/R526V/T560W | Aeropyrum pernix |